Cell adhesion inhibitors

ABSTRACT

The present invention relates to novel compounds that are useful for inhibition and prevention of cell adhesion and cell adhesion-mediated pathologies. This invention also relates to pharmaceutical formulations comprising these compounds and methods of using them for inhibition and prevention of cell adhesion and cell adhesion-mediated pathologies. The compounds and pharmaceutical compositions of this invention can be used as therapeutic or prophylactic agents. They are particularly well-suited for treatment of many inflammatory and autoimmune diseases.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.08/983,391, filed on Aug. 10, 1998 now U.S. Pat. No. 6,239,108, whichclaims priority from PCT/US96/11570 filed on Jul. 11, 1996, and fromU.S. application Ser. No. 08/498,237, filed on Jul. 11, 1995, each ofwhich is incorporated by reference in its entirety.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to novel compounds that are useful forinhibition and prevention of cell adhesion and cell adhesion-mediatedpathologies. This invention also relates to pharmaceutical formulationscomprising these compounds and methods of using them for inhibition andprevention of cell adhesion and cell adhesion-mediated pathologies. Thecompounds and pharmaceutical compositions of this invention can be usedas therapeutic or prophylactic agents. They are particularly well-suitedfor treatment of many inflammatory and autoimmune diseases.

BACKGROUND OF THE INVENTION

Cell adhesion is a process by which cells associate with each other,migrate towards a specific target or localize within the extra-cellularmatrix. As such, cell adhesion constitutes one of the fundamentalmechanisms underlying numerous biological phenomena. For example, celladhesion is responsible for the adhesion of hematopoietic cells toendothelial cells and the subsequent migration of those hemopoieticcells out of blood vessels and to the site of injury. As such, celladhesion plays a role in pathologies such as inflammation and immunereactions in mammals.

Investigations into the molecular basis for cell adhesion have revealedthat various cell-surface macromolecules—collectively known as celladhesion molecules or receptors—mediate cell-cell and cell-matrixinteractions. For example, proteins of the superfamily called“integrins” are key mediators in adhesive interactions betweenhematopoietic cells and their microenvironment (M. E. Hemler, “VLAProteins in the Integrin Family: Structures, Functions, and Their Roleon Leukocytes.”, Ann. Rev. Immunol., 8, p. 365 (1990)). Integrins arenon-covalent heterodimeric complexes consisting of two subunits called αand β. There are at least 12 different α subunits α1, α2, α3, α4, α5,α6, αL, αM, αX, αIIB, αV, and αE) and at least 9 different β (β1-β9)subunits. Based on the type of its α and β subunit components, eachintegrin molecule is categorized into a subfamily.

α4β1 integrin, also known as very late antigen-4 (“VLA-4”) orCD49d/CD29, is a leukocyte cell surface receptor that participates in awide variety of both cell-cell and cell-matrix adhesive interactions (M.E. Hemler, Ann. Rev. Immunol., 8, p. 365 (1990)). It serves as areceptor for the cytokine-inducible endothelial cell surface protein,vascular cell adhesion molecule-1 (“VCAM-1”), as well as to theextracellular matrix protein fibronectin (“FN”) (Ruegg et al., J. CellBiol., 177, p. 179 (1991); Wayner et al., J. Cell Biol., 105, p. 1873(1987); Kramer et al., J. Biol. Chem., 264, p. 4684 (1989); Gehlsen etal. Science, 24, p. 1228 (1988)). Anti-VLA4 monoclonal antibodies(“mAb's”) have been shown to inhibit VLA4-dependent adhesiveinteractions both in vitro and in vivo (Ferguson et al. Proc. Natl.Acad. Sci., 88, p. 8072 (1991); Ferguson et al., J. Immunol., 150, p.1172 (1993)). Results of in vivo experiments suggest that thisinhibition of VLA-4-dependent cell adhesion may prevent or inhibitseveral inflammatory and autoimmune pathologies (R. L. Lobb et al., “ThePathophysiologic Role of α4 Integrins In Vivo”, J. Clin. Invest., 94,pp. 1722-28 (1994)).

In order to identify the minimum active amino acid sequence necessary tobind VLA-4, Komoriya et al. (“The Minimal Essential Sequence for a MajorCell Type-Specific Adhesion Site (CS1) Within the Alternatively SplicedType III Connecting Segment Domain of Fibronectin Is Leucine-AsparticAcid-Valine”, J. Biol. Chem., 266 (23), pp. 15075-79 (1991)) synthesizeda variety of overlapping peptides based on the amino acid sequence ofthe CS-1 region (the VLA-4 binding domain) of a particular species offibronectin. They identified an 8-amino acid peptide,Glu-Ile-Leu-Asp-Val-Pro-Ser-Thr [SEQ ID NO: 1], as well as two smalleroverlapping pentapeptides, Glu-Ile-Leu-Asp-Val [SEQ ID NO: 2] andLeu-Asp-Val-Pro-Ser [SEQ ID NO: 3], that possessed inhibitory activityagainst FN-dependent cell adhesion. These results suggested thetripeptide Leu-Asp-Val as a minimum sequence for cell-adhesion activity.It was later shown that Leu-Asp-Val binds only to lymphocytes thatexpress an activated form of VLA-4, thus bringing into question theutility of such a peptide in vivo (E. A. Wayner et al.,“Activation-Dependent Recognition by Hematopoietic Cells of the LDVSequence in the V Region of Fibronectin”, J. Cell. Biol., 116(2), pp.489-497 (1992)). However, certain larger peptides containing the LDVsequence were subsequently shown to be active in vivo [T. A. Ferguson etal., “Two Integrin Binding Peptides Abrogate T-cell-Mediated ImmuneResponses In Vivo”, Proc. Natl. Acad. Sci. USA, 88, pp. 8072-76 (1991);and S. M. Wahl et al., “Synthetic Fibronectin Peptides SuppressArthritis in Rats by Interrupting Leukocyte Adhesion and Recruitment”,J. Clin. Invest., 94, pp. 655-62 (l994)].

A cyclic pentapeptide, Arg-Cys-Asp-TPro-Cys (wherein TPro denotes4-thioproline) (SEQ ID NO:5), which can inhibit both VLA-4 and VLA-5adhesion to FN has also been described (D. M. Nowlin et al. “A NovelCyclic Pentapeptide Inhibits α4β1 and α5β1 Integrin-mediated CellAdhesion”, J. Biol. Chem., 268(27), pp. 20352-59 (1993); and PCTpublication PCT/US91/04862). This peptide was based on the tripeptuidesequence Arg-Gly-Asp from FN which had been known as a common motif inthe recognition site for several extracellular-matrix proteins.

Despite these advances, there remains a need for small, specificinhibitors of VLA-4-dependent cell adhesion. Ideally, such inhibitorswould be semi-peptidic or non-peptidic so that they may be orallyadministered. Such compounds would provide useful agents for treatment,prevention or suppression of various pathologies mediated by celladhesion and VLA-4 binding. co-pending/United States patent application08/376,372 describes β-amino acid containing linear peptidyl compoundswith cell adhesion inhibitory activity. International patentapplications WO 94/15958 and WO 92/00995 describe cyclic peptide andpeptidomimetic compounds with cell adhesion modulating activity.International patent applications WO 93/08823 and WO 92/08464 describeguanidinyl-, urea- and thiourea-containing cell adhesion modulatingcompounds. U.S. Pat. No. 5,260,277 describes guanidinyl cell adhesionmodulation compounds.

SUMMARY OF THE INVENTION

The present invention solves this problem by providing novelsemi-peptidic compounds that inhibit the binding of ligands to VLA-4.These compounds are useful for inhibition, prevention and suppression ofVLA-4-mediated cell adhesion and pathologies associated with thatadhesion, such as inflammation and immune reactions. The compounds ofthis invention may be used alone or in combination with othertherapeutic or prophylactic agents to inhibit, prevent or suppress celladhesion. This invention also provides pharmaceutical formulationscontaining these VLA-4-mediated cell adhesion inhibitors and methods ofusing the compounds and compositions of the invention for inhibition ofcell adhesion.

According to one embodiment of this invention, these novel compounds,compositions and methods are advantageously used to treat inflammatoryand immune diseases. The present invention also provides methods forpreparing the compounds of this invention and intermediates useful inthose methods.

DETAILED DESCRIPTION OF THE INVENTION

The following abbreviations are used in the description:

Designation Reagent or Fragment Ac acetyl Bn benzyl Boctert-butoxycarbonyl Bu butyl Cbz carbobenzyloxy Cy cyclohexyl CyMcyclohexylmethyl DIPEA diisopropylethylamine EDC1-(3-diethylaminopropyl)-3- ethylcarbodiimide HOBT1-hydroxybenzotriazole hydrate i-amyl isoamyl i-Pn isopentyl i-Prisopropyl Me methyl 2-MPUBA 4-(N′-(2-methylphenyl)urea)-phenylmethylamino 2-MPUPA 4-(N′-(2-methylphenyl)urea)- phenylacetyl NMPN-methylpyrrolidinone NMM N-methylmorpholine Ph phenyl PUPA4-(N′-phenylurea)phenylacetyl Su succinimidyl TBTU2-(1H-benzotriazol-1-yl)- 1,1,3,3-tetramethyluronium tetrafluoroborateTEA triethylamine TFA trifluoroacetic acid THAMtris(hydroxy)methylaminomethane

Definitions

As used herein, the term “alkyl”, alone or in combination, refers to astraight-chain or branched-chain alkyl radical containing from 1 to 10,preferably from 1 to 6 and more preferably from 1 to 4, carbon atoms.Examples of such radicals include, but are not limited to, methyl,ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl,pentyl, iso-amyl, hexyl, decyl and the like.

The term “alkenyl”, alone or in combination, refers to a straight-chainor branched-chain alkenyl radical containing from 2 to 10, preferablyfrom 2 to 6 and more preferably from 2 to 4, carbon atoms. Examples ofsuch radicals include, but are not limited to, ethenyl, E- andZ-propenyl, isopropenyl, E- and Z-butenyl, E- and Z-isobutenyl, E- andZ-pentenyl, decenyl and the like.

The term “alkynyl”, alone or in combination, refers to a straight-chainor branched-chain alkynyl radical containing from 2 to 10, preferablyfrom 2 to 6 and more preferably from 2 to 4, carbon atoms. Examples ofsuch radicals include, but are not limited to, ethynyl (acetylenyl),propynyl, propargyl, butynyl, hexynyl, decynyl and the like.

The term “cycloalkyl”, alone or in combination, refers to a cyclic alkylradical containing from 3-10, preferably from 3-8 and more preferablyfrom 3-6, carbon atoms and may be optionally aryl-fused. Examples ofsuch radicals include, but are not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl and the like.

The term “cycloalkenyl”, alone or in combination, refers to a cycliccarbocycle containing from 4 to 8, preferably 5 or 6, carbon atoms andone or more double bonds. Examples of such cycloalkenyl radicalsinclude, but are not limited to, cyclopentenyl, cyclohexenyl,cyclopentadienyl and the like.

The term “aryl” refers to a carbocyclic aromatic group selected from thegroup consisting of phenyl, naphthyl, indenyl, indanyl, azulenyl,fluorenyl, and anthracenyl; or a heterocyclic aromatic group selectedfrom the group consisting of furyl, thienyl, pyridyl, pyrrolyl,oxazolyly, thiazolyl, imidazolyl, pyrazolyl, 2-pyrazolinyl,pyrazolidinyl, isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl,1,2,3-triazolyl, 1,3,4-thiadiazolyl, pyridazinyl, pyrimidinyl,pyrazinyl, 1,3,5-triazinyl, 1,3,5-trithianyl, indolizinyl, indolyl,isoindolyl, 3H-indolyl, indolinyl, benzo[b]furanyl,2,3-dihydrobenzofuranyl, benzo[b]thiophenyl, 1H-indazolyl,benzimidazolyl, benzthiazolyl, purinyl, 4H-quinolizinyl, quinolinyl,isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl,1,8-naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl,phenothiazinyl, phenoxazinyl, pyrazolo[1,5-c]triazinyl and the like.

“Aryl”, “cycloalkyl” and “cycloalkenyl” groups, as defined in thisapplication may independently contain up to three substituents which areindependently selected from the group consisting of halogen, hydroxyl,amino, nitro, trifluoromethyl, trifluoromethoxy, alkyl, alkenyl,alkynyl, cyano, carboxy, carboalkoxy, Ar′-substituted alkyl,Ar′-substituted alkenyl or alkynyl, 1,2-dioxymethylene,1,2-dioxyethylene, alkoxy, alkenoxy or alkynoxy, Ar′-substituted alkoxy,Ar′-substituted alkenoxy or alkynoxy, alkylamino, alkenylamino oralkynylamino, Ar′-substituted alkylamino, Ar′-substituted alkenylaminoor alkynylamino, Ar′-substituted carbonyloxy, alkylcarbonyloxy,aliphatic or aromatic acyl, Ar′-substituted acyl, Ar′-substitutedalkylcarbonyloxy, Ar′-substituted carbonylamino, Ar′-substituted amino,Ar′-substituted oxy, Ar′-substituted carbonyl, alkylcarbonylamino,Ar′-substituted alkylcarbonylamino, alkoxy-carbonylamino,Ar′-substituted alkoxycarbonyl-amino, Ar′-oxycarbonylamino,alkylsulfonylamino, mono- or bis-(Ar′-sulfonyl)amino, Ar′-substitutedalkyl-sulfonylamino, morpholinocarbonylamino,thiomorpholinocarbonylamino, N-alkyl guanidino, N-Ar′ guanidino,N-N-(Ar′,alkyl) guanidino, N,N-(Ar′,Ar′)guanidino, N,N-dialkylguanidino, N,N,N-trialkyl guanidino, N-alkyl urea, N,N-dialkyl urea,N-Ar′ urea, N,N-(Ar′,alkyl) urea, N,N-(Ar′)₂ urea,aralkyloxycarbonyl-substituted alkyl, aralkylaminocarbonyl, thioaryloxyand the like; wherein “Ar′” is defined similarly to aryl, but containsup to three substituents selected from the group consisting of halogen,hydroxyl, amino, nitro, trifluoromethyl, trifluoromethoxy, alkyl,alkenyl, alkynyl, 1,2-dioxymethylene, 1,2-dioxyethylene, alkoxy,alkenoxy, alkynoxy, alkylamino, alkenylamino or alkynylamino,alkylcarbonyloxy, aliphatic or aromatic acyl, alkylcarbonylamino,alkoxycarbonylamino, alkylsulfonylamino, N-alkyl or N,N-dialkyl urea.

The term “aralkyl”, alone or in combination, refers to an arylsubstituted alkyl radical, wherein the term “alkyl” and “aryl” are asdefined above. Examples of suitable aralkyl radicals include, but arenot limited to, phenylmethyl, phenethyl, phenylhexyl, diphenylmethyl,pyridylmethyl, tetrazolylmethyl, furylmethyl, imidazolylmethyl,indolylmethyl, thienylpropyl and the like.

The term “alkoxy”, alone or in combination, refers to an alkyl etherradical, wherein the term “alkyl” is as defined above. Examples ofsuitable alkyl ether radicals include, but are not limited to, methoxy,ethoxy, n-propoxy, iso-propoxy, n-butoxy, iso-butoxy, sec-butoxy,tert-butoxy and the like.

The term “alkenoxy”, alone or in combination, refers to a radical offormula alkenyl-O—, wherein the term “alkenyl” is as defined aboveprovided that the radical is not an enol ether. Examples of suitablealkenoxy radicals include, but are not limited to, allyloxy, E- andZ-3-methyl-2-propenoxy and the like.

The term “alkynyloxy”, alone or in combination, refers to a radical offormula alkynyl-O—, wherein the term “alkynyl” is as defined aboveprovided that the radical is not an ynol ether. Examples of suitablealkynoxy radicals include, but are not limited to, propargyloxy,2-butynyloxy and the like.

The term “thioalkoxy” refers to a thioether radical of formula alkyl-S—,wherein alkyl is as defined above.

The term “alkylamino”, alone or in combination, refers to a mono- ordi-alkyl-substituted amino radical (i.e., a radical of formula alkyl-NH—or (alkyl)₂-N—), wherein the term “alkyl” is as defined above. Examplesof suitable alkylamino radicals include, but are not limited to,methylamino, ethylamino, propylamino, isopropylamino, t-butylamino,N,N-diethylamino and the like.

The term “alkenylamino”, alone or in combination, refers to a radical offormula alkenyl-NH— or (alkenyl)₂N—, wherein the term “alkenyl” is asdefined above, provided that the radical is not an enamine. An exampleof such alkenylamino radicals is the allylamino radical.

The term “alkynylamino”, alone or in combination, refers to a radical offormula alkynyl-NH— or (alkynyl)₂N—, wherein the term “alkynyl” is asdefined above, provided that the radical is not an ynamine. An exampleof such alkynylamino radicals is the propargyl amino radical.

The term “aryloxy”, alone or in combination, refers to a radical offormula aryl-O—, wherein aryl is as defined above. Examples of aryloxyradicals include, but are not limited to, phenoxy, naphthoxy, pyridyloxyand the like.

The term “arylamino”, alone or in combination, refers to a radical offormula aryl-NH—, wherein aryl is as defined above. Examples ofarylamino radicals include, but are not limited to, phenylamino(anilido), naphthylamino, 2-, 3- and 4-pyridylamino and the like.

The term “biaryl”, alone or in combination, refers to a radical offormula aryl-aryl-, wherein the term “aryl” is as defined above.

The term “thioaryl”, alone or in combination, refers to a radical offormula aryl-S—, wherein the term “aryl” is as defined above. An exampleof a thioaryl radical is the thiophenyl radical.

The term “aryl-fused cycloalkyl”, alone or in combination, refers to acycloalkyl radical which shares two adjacent atoms with an aryl radical,wherein the terms “cycloalkyl” and “aryl” are as defined above. Anexample of an aryl-fused cycloalkyl radical is the benzo-fusedcyclobutyl radical.

The term “aliphatic acyl”, alone or in combination, refers to radicalsof formula alkyl-CO—, alkenyl-CO— and alkynyl-CO— derived from analkane-, alkene- or alkyncarboxylic acid, wherein the terms “alkyl”,“alkenyl” and “alkynyl” are as defined above. Examples of such aliphaticacyl radicals include, but are not limited to, acetyl, propionyl,butyryl, valeryl, 4-methylvaleryl, acryloyl, crotyl, propiolyl,methylpropiolyl and the like.

The terms “aromatic acyl” or “aroyl”, alone or in combination, refers toa radical of formula aryl-CO—, wherein the term “aryl” is as definedabove. Examples of suitable aromatic acyl radicals include, but are notlimited to, benzoyl, 4-halobenzoyl, 4-carboxybenzoyl, naphthoyl,pyridylcarbonyl and the like.

The term “heterocycloyl”, alone or in combination, refers to radicals offormula heterocycle-CO—, wherein the term “heterocycle” is as definedbelow. Examples of suitable heterocycloyl radicals include but are notlimited to, tetrahydrofuranylcarbonyl, piperidinylcarbonyl,tetrahydrothiophenecarbonyl and the like.

The terms “morpholinocarbonyl” and “thiomorpholinocarbonyl”, alone or incombination with other terms, refer to an N-carbonylated morpholino andan N-carbonylated thiomorpholino radical, respectively.

The term “alkylcarbonylamino”, alone or in combination, refers to aradical of formula alkyl-CONH, wherein the term “alkyl” is as definedabove.

The term “alkoxycarbonylamino”, alone or in combination, refers to aradical of formula alkyl-OCONH—, wherein the term “alkyl” is as definedabove.

The term “alkylsulfonylamino”, alone or in combination, refers to aradical of formula alkyl-SO₂NH—, wherein the term “alkyl” is as definedabove.

The term “arylsulfonylamino”, alone or in combination, refers to aradical of formula aryl-SO₂NH—, wherein the term “aryl” is as definedabove.

The term “N-alkylurea”, alone or in combination, refers to a radical offormula alkyl-NH—CO—NH—, wherein the term “alkyl” is as defined above.

The term “N-arylurea”, alone or in combination, refers to a radical offormula aryl-NH—CO—NH—, wherein the term “aryl” is as defined above.

The term “halogen” means fluorine, chlorine, bromine and iodine.

The terms “heterocycle” and “heterocyclic ring”, alone or incombination, refer to a non-aromatic 3- to 10-membered ring containingat least one endocyclic N, O or S atom. The heterocycle may optionallybe aryl-fused. The heterocycle may also be optionally substituted withone to three substituents which are independently selected from thegroup consisting of hydrogen, halogen, hydroxyl, amino, nitro,trifluoromethyl, trifluoromethoxy, alkyl, aralkyl, alkenyl, alkynyl,aryl, cyano, carboxy, carboalkoxy, Ar′-substituted alkyl,Ar′-substituted alkenyl or alkynyl, 1,2-dioxymethylene,1,2-dioxyethylene, alkoxy, alkenoxy or alkynoxy, Ar′-substituted alkoxy,Ar′-substituted alkenoxy or alkynoxy, alkylamino, alkenylamino oralkynylamino, Ar′-substituted alkylamino, Ar′-substituted alkenylaminoor alkynylamino, Ar′-substituted carbonyloxy, alkylcarbonyloxy,aliphatic or aromatic acyl, Ar′-substituted acyl, Ar′-substitutedalkylcarbonyloxy, Ar′-substituted carbonylamino, Ar′-substituted amino,Ar′-substituted oxy, Ar′-substituted carbonyl, alkylcarbonylamino,Ar′-substituted alkylcarbonylamino, alkoxy-carbonylamino,Ar′-substituted alkoxycarbonyl-amino, Ar′-oxycarbonylamino,alkylsulfonylamino, mono- or bis-(Ar′-sulfonyl)amino, Ar′-substitutedalkyl-sulfonylamino, morpholinocarbonylamino,thiomorpholinocarbonylamino, N-alkyl guanidino, N-Ar′ guanidino,N-N-(Ar′,alkyl) guanidino, N,N-(Ar′,Ar′)guanidino, N,N-dialkylguanidino, N,N,N-trialkyl guanidino, N-alkyl urea, N,N-dialkyl urea,N-Ar′ urea, N,N-(Ar′,alkyl) urea, N,N-(Ar′)₂ urea,aralkoxycarbonyl-substituted alkyl, carboxyalkyl, oxo, arylsulfonyl andaralkylaminocarbonyl.

The term “leaving group” generally refers to groups readily displaceableby a nucleophile, such as an amine, and alcohol or a thiol nucleophile.Such leaving groups are well known and include carboxylates,N-hydroxysuccinimide, N-hydroxybenzotriazole, halogen (halides),triflates, tosylates, mesylates, alkoxy, thioalkoxy and the like.

The term “hydrophobic group” refers to a group which is resistant touniting with or absorbing water. Examples of such hydrophobic groupsinclude, but are not limited to, methyl, ethyl, propy, butyl, pentyl,hexyl, phenyl, benzyl, naphthyl, N-benzylimidazolyl, methylthioethyl andthe like.

The term “acidic functional group” refers to a group which has an acidichydrogen within it. Examples of such groups include, but are not limitedto, carboxylic acid, tetrazole, imidazole, hydroxyl, mercapto,hydroxylaminocarbonyl, sulfonic acid, sulfinic acid, phosphoric acid andphosphonic acid.

The terms “activated derivative of a suitably protected α-amino acid”and “activated substituted-phenylacetic acid derivative” refer toderivatives of carboxylic acids wherein the —OH group is replaced by asuperior leaving group. Examples of activated acid derivatives include,but are not limited to, the corresponding acyl halides (e.g. acidfluoride, acid chloride and acid bromide), corresponding activatedesters (e.g. nitrophenyl ester, the ester of 1-hydroxybenzotriazole,HOBT, or the ester of hydroxysuccinimide, HOSu), and other conventionalderivatives within the skill of the art.

The terms “protected” or “protecting group” refer to a suitable chemicalgroup which may be attached to a functional group of a molecule, thenremoved at a later stage to reveal the intact functional group andmolecule. Examples of suitable protecting groups for various functionalgroups are described in T. W. Greene and P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, 2d. Ed., John Wiley and Sons (1991); L.Fieser and M. Fieser, Fieser and Fieser's Reagents for OrganicSynthesis, John Wiley and Sons (1994); L. Paquette, ed. Encyclopedia ofReagents for Organic Synthesis, John Wiley and Sons (1995).

The compounds of this invention contain one or more asymmetric carbonatoms and thus occur as racemates and racemic mixtures, singleenantiomers, diastereomeric mixtures and individual diastereomers. Allsuch isomeric forms of these compounds are expressly included in thepresent invention. Each stereogenic carbon may be of the R or Sconfiguration. Although the specific compounds exemplified in thisapplication may be depicted in a particular stereochemicalconfiguration, compounds having either the opposite stereochemistry atany given chiral center or mixtures thereof are envisioned as part ofthe invention. Although amino acids and amino acid side chains may bedepicted in a particular configuration, both natural and unnatural formsare envisioned as part of the invention.

In view of the above definitions, other chemical terms used throughoutthis application can be easily understood by those of skill in the art.Terms may be used alone or in any combination thereof. The preferred andmore preferred chain lengths of the radicals apply to all suchcombinations.

This invention provides compounds which are capable of inhibitingVLA-4-mediated cell adhesion by inhibiting the binding of ligands tothat receptor. These compounds are represented by formula (I):Z—(Y¹)—(Y²)—(Y³)—X  (I)and pharmaceutically acceptable derivatives thereof; wherein:

Z is selected from the group consisting of alkyl; aliphatic acyloptionally substituted with N-alkyl- or N-arylamido; aroyl;heterocycloyl; alkyl- or arylsulfonyl; aralkylcarbonyl optionallysubstituted with aryl; heterocycloalkylcarbonyl; alkoxycarbonyl;aralkyloxycarbonyl; cycloalkylcarbonyl optionally fused with aryl;heterocycloalkoxycarbonyl; alkylaminocarbonyl; arylamino carbonyl andaralkylaminocarbonyl optionally substituted withbis(alkylsulfonyl)amino, alkoxycarbonylamino or alkenyl; alkylsulfonyl;aralkylsulfonyl; arylsulfonyl; cycloalkylsulfonyl optionally fused witharyl; heterocyclylsulfonyl; heterocyclylalkylsulfonyl; aralkoxycarbonyl;aryloxycarbonyl; cycloalkyloxycarbonyl; heterocyclyloxycarbonyl;heterocyclylalkoxycarbonyl; mono- or di-alkylaminocarbonyl optionallysubstituted with aryl; (alkyl) (aralkyl)aminocarbonyl; mono- ordi-aralkylaminocarbonyl; mono- or di-arylaminocarbonyl; (aryl)(alkyl)aminocarbonyl; mono- or di-cycloalkylaminocarbonyl;heterocyclylaminocarbonyl; heterocyclylalkylaminocarbonyl; (alkyl)(heterocyclyl)aminocarbonyl; (alkyl) (heterocyclylalkyl)aminocarbonyl;(aralkyl) (heterocyclyl)aminocarbonyl; (aralkyl)(heterocyclylalkyl)aminocarbonyl; alkenoyl optionally substituted witharyl; alkenylsulfonyl optionally substituted with aryl; alkynoyloptionally substituted with aryl; alkynylsulfonyl optionally substitutedwith aryl; cycloalkenylcarbonyl; cycloalkenylsulfonyl;cycloalkylalkanoyl; cycloalkylalkylsulfonyl; arylaroyl, biarylsulfonyl;alkoxysulfonyl; aralkoxysulfonyl; alkylaminosulfonyl; aryloxysulfonyl;arylaminosulfonyl; N-arylurea-substituted alkanoyl;N-arylurea-substituted alkylsulfonyl; cycloalkenyl-substituted carbonyl;cycloalkenyl-substituted sulfonyl; alkenoxycarbonyl optionallysubstituted with aryl; alkenoxysulfonyl optionally substituted witharyl; alkynoxycarbonyl optionally substituted with aryl;alkynoxysulfonyl optionally substituted with aryl; alkenyl- oralkynyl-aminocarbonyl optionally substituted with aryl; alkenyl- oralkynyl-aminosulfonyl optionally substituted with aryl;acylamino-substituted alkanoyl; acylamino-substituted alkylsulfonyl;aminocarbonyl-substituted alkanoyl; carbamoyl-substituted alkanoyl;carbamoyl-substituted alkylsulfonyl; heterocyclylalkanoyl;heterocyclylaminosulfonyl; carboxyalkyl-substituted aralkoyl;carboxyalkyl-substituted aralkylsulfonyl; oxocarbocyclyl-fused aroyl;oxocarbocyclyl-fused arylsulfonyl; heterocyclylalkanoyl; N′,N′-alkyl,arylhydrazinocarbonyl; aryloxy-substituted alkanoyl andheterocyclylalkylsulfonyl.

Y¹ is —N(R¹)—C(R²)(A¹)—C(O)—;

Y² is —N(R¹)—C(R²)(A²)—C(O)—;

each Y³ is represented by the formula —N(R¹)—C(R²)(A³)—C(O)—;

each R¹ is independently selected from the group consisting of hydrogen,alkyl, and aralkyl; alkenyl; alkynyl; cycloalkyl; cycloalkenyl;cycloalkylalkyl; aryl; aminoalkyl; mono- or di-alkyl-substitutedaminoalkyl; mono- or di-aralkyl-substituted aminoalkyl; hydroxyalkyl;alkoxyalkyl; mercaptoalkyl; thioalkoxyalkyl

A¹ is selected from the group consisting of amino acid side chains andcorresponding protected derivatives; cycloalkyl; and alkyl optionallysubstituted with amino, acylamino, amino-substituted acylamino,alkoxycarbonylamino, aryl, cycloalkyl, carboxy, alkoxy, aralkyloxy,alkoxycarbonyl, aralkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, (alkyl) (aralkyl)aminocarbonyl,aralkylaminocarbonyl, diaralkylaminocarbonyl, hydroxyl,carboxyalkylaminocarbonyl, hydroxylaminocarbonyl, mercapto, thioalkoxyor heterocycle;

A² is selected from the group consisting of acidic functional groups andalkyl optionally substituted with an acidic functional group, protectedacidic functional group or aryl;

each A³ is independently selected from the group consisting of aminoacid side chains and corresponding protected derivatives; aryl;cycloalkyl; and alkyl optionally substituted with amino, acylamino,amino-substituted acylamino, aryl, cycloalkyl, carboxy, alkoxy,aralkyloxy, alkoxycarbonyl, aralkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, (alkyl)(aralkyl)aminocarbonyl, aralkylaminocarbonyl, diaralkylaminocarbonyl,hydroxyl, carboxyalkylaminocarbonyl, hydroxylaminocarbonyl, mercapto,thioalkoxy or heterocycle;

or R¹ and any A are taken together with the atoms to which they areattached form a 3- to 6-membered ring heterocycle;

each R² is independently selected from the group consisting of hydrogenand alkyl;

n is an integer from 0 to 8; and

X is selected from the group consisting of alkoxy; aryloxy; aralkyloxy;hydroxyl; amino; alkylamino optionally substituted with hydroxy,aminocarbonyl, N-alkylaminocarbonyl, carboxy or alkoxycarbonyl;dialkylamino; cycloalkylamino; dicycloalkylamino; cycloalkylalkylamino;(alkyl) (aryl)amino; aralkylamino optionally substituted with carboxy;diaralkylamino; arylamino; heterocycle; and (mono- or bis-carboxylicacid)-substituted alkylamine; heterocyclylamino;heterocyclyl-substituted alkylamino and wherein the compound of formulaI is expressly notN′-carboxymethyl-N-(phenylacetyl-L-leucyl-L-aspartyl-L-phenylalanyl-L-prolyl)piperazine(i.e., when Z=phenylacetyl, Y¹=L, Y²=D, Y³=F/P, n=2, andX=4-carboxymethylpiperazinyl) and expressly notphenylacetyl-L-leucyl-L-aspartyl-L-phenylalanyl-D-proline amide (i.e.,when Z=phenylacetyl, Y¹=L, y²=D, Y³=F/p, n=2, and X=NH₂).

A “pharmaceutically acceptable derivative” denotes any pharmaceuticallyacceptable salt, ester, salt of such ester, amide or salt of such amideof a compound of this invention. The invention also includes any othercompound which, upon administration to a patient, is capable ofproviding (directly or indirectly) a compound of this invention (e.g. aprodrug). The invention also includes metabolites or residues of acompound of this invention characterized by the ability to inhibit,prevent or suppress cell adhesion and cell adhesion-mediatedpathologies.

In a preferred embodiment of this invention, Al is selected from thegroup consisting of cycloalkyl; heterocyclic ring (when A¹ and R¹ aretaken together); and alkyl optionally substituted with amino, acylamino,amino-substituted acylamino, aryl, carboxy, cycloalkyl, hydroxy, alkoxy,aralkyloxy, alkoxycarbonyl, aralkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, (alkyl)(aralkyl)aminocarbonyl,aralkylaminocarbonyl, diaralkylaminocarbonyl, alkoxycarbonylamino,mercapto, thioalkoxy or heterocycle.

More preferably, A¹ is selected from the group consisting ofaminocarbonylethyl, benzyl, n-butyl, isobutyl, carboxyethyl, cyclohexyl,1-hydroxyethyl, hydroxymethyl, mercaptomethyl, 1-methylpropyl,methylthioethyl, n-propyl, isopropyl, methoxycarbonylaminobutyl,6-aminohexanoylaminobutyl and (when A¹ and R¹ are taken together)azetidine, aziridine, pyrrolidine, and piperidine.

Even more preferably, A¹ is selected from the group consisting ofbenzyl, n-butyl, isobutyl, methylthioethyl, cyclohexyl, 1-methylpropyl,n-propyl and isopropyl. An alternate preferred A¹ is (when A¹ and R¹ aretaken together) pyrrolidine.

In an alternate preferred embodiment, A² is selected from the groupconsisting of alkyl optionally substituted with amino, aminocarbonyl,aryl, alkoxycarbonyl, aralkyloxycarbonyl, hydroxylaminocarbonyl,carboxy, NH-containing heterocycle, hydroxy, or mercapto; aralkyloptionally substituted with amino, aminocarbonyl, carboxy, NH-containingheterocycle, hydroxy, or mercapto; and heterocyclic ring (when A² and R¹are taken together).

More preferably, A² is selected from the group consisting ofcarboxymethyl, 2-carboxyethyl, 1-carboxyethyl,hydroxylaminocarbonylmethyl, hydroxymethyl, mercaptomethyl,imidazolylmethyl, N-Bn-imidazolylmethyl, phenyl, carbomethoxymethyl,carbobenzyloxymethyl, and (when A² and R¹ are taken together) azetidine,aziridine, pyrrolidine, and piperidine.

Even more preferably, A² is selected from the group consisting ofcarboxymethyl, 2-carboxyethyl, 1-carboxyethyl,hydroxylaminocarbonylmethyl, hydroxymethyl, mercaptomethyl andimidazolylmethyl.

According to another preferred embodiment, A³ is independently selectedfrom the group consisting of amino acid side chains and correspondingprotected derivatives; cycloalkyl; and alkyl optionally substituted witharyl, cycloalkyl, carboxy, hydroxylaminocarbonyl, alkoxy, aralkyloxy,mercapto, N-containing heterocycle, carboxyalkylaminocarbonyl oramino-substituted acylamino.

More preferably, A³ is independently selected from the group consistingof amino acid side chains and corresponding protected derivatives;cyclohexyl; and alkyl optionally substituted with phenyl, cyclohexyl,carboxy, hydroxylaminocarbonyl, methoxy, benzyloxy, mercapto,N-benzylimidazolyl, biotinyl, tetrazolyl, valinyl-N-carbonyl or6-aminohexanoylamino.

According to another preferred embodiment, each Y³ is independentlyselected from the group consisting of amino acids and correspondingprotected derivatives.

According to another preferred embodiment, Y¹ is leucinyl (R¹=H, R²=H,A¹=i-Bu); Y² is aspartyl (R¹=H, R²=H, A²=carboxymethyl); n=2; and Y³ isvalinylprolinyl (R¹=H, R²=H, A³=i-Pr)/(R²=H, R¹ with A³=proline).

In another preferred embodiment, X is selected from the group consistingof alkoxy; aryloxy; aralkyloxy; hydroxyl; amino; mono- and dialkylaminooptionally substituted with hydroxy, aminocarbonyl,N-alkylaminocarbonyl, carboxy or alkoxycarbonyl; dialkylamino;cycloalkylamino; cycloalkylalkylamino; dicycloalkylamino;(alkyl)(aryl)amino; aralkylamino optionally substituted with carboxy;diaralkylamino; arylamino; N-containing heterocycle; bis-carboxylicacid-substituted alkylamine and (mono- orbis-carboxy)methylaminocarbonyl-substituted-N-containing heterocycle.

More preferably, X is selected from the group consisting of amino,methylamino, isopropylamino, isobutylamino, n-butylamino, t-butylamino,isoamylamino, isopentylamino, hexylamino, cyclohexylamino,cyclohexylmethylamino, methylphenylamino, phenylmethylamino,phenylamino, 4-methoxyphenylmethylamino, dimethylamino,diisopropylamino, diisobutylamino, hydroxy, methoxy, n-butoxy, t-butoxy,benzyloxy, 2-piperidinecarboxylic acid,N′-(α,α′-bis-carboxymethyl)-2-piperidinecarboxamide,N′-carboxymethyl-2-piperidinecarboxamide,1-hydroxymethyl-2-methylpropylamino,1-N′-methylamido-1-methylethylamino, 3,3-dimethylbutylamino,1-N′-methylamidobutylamino, 1-amido-2-methylbutylamino,1-carbomethoxy-2-methylbutylamino, 1-N′-methylamido-2-methylbutylamino,1-carboxy-1-phenylmethylamino, morpholino, piperidinyl,N-phenylpiperazinyl, pipecolinyl, and piperazinyl.

According to another preferred embodiment, Z is selected from the groupconsisting of aliphatic acyl, aroyl, aralkylcarbonyl, heterocycloyl,alkoxycarbonyl, aralkyloxycarbonyl and heterocycloalkylcarbonyl. Morepreferably, Z is a (N-Ar′-urea)-para-substituted aralkylcarbonyl groupand even more preferably, Z is a (N-Ar′-urea)-para-substitutedphenylmethylcarbonyl group or (N-Ar′-urea)-para-substitutedpyridylmethylcarbonyl group. Even more preferably, Z is a(N-ortho-substituted-Ar′urea)-para-substituted phenylmethylcarbonylgroup or (N-meta-substituted-Ar′urea)-para-substitutedphenylmethylcarbonyl group.

Examples of some specific preferred compounds of this invention areprovided in Table 1.

TABLE 1 Z-(Y¹)—(Y²)—(Y³)—X (I) wherein Y¹ is —N(R¹)—C(R²) (A¹)-C(O)—; Y²is —N(R¹)—C(R²) (A²)-C(O)—; each Y³ is represented by the formula—N(R¹)—C(R²) (A³)-C(O)—; For A¹, A² and A³, a single letter code refersto the side chain of the corresponding amino acid designated by thatletter. A capital letter (e.q., A) indicates the L-amino acid while asmall letter (e.g., a) indicates the D-amino acid. Both capital andsmall letters (e.g., L(l)) indicates a mixture. Unless expressly notedto the contrary; compounds in this table have R¹ and R² as hydrogen.Cmpd # Z A¹ A² (A³⁾n X  1 3-methoxy-4-(N′-phenyl L D V/P OHurea)phenylacetyl  2 3-methoxy-4-(N′-phenyl M D V/P OH urea)phenylacetyl 3 6-methoxy-5-(N′-(2-methylphenyl)- L D V NH₂ urea)-2-pyridylacetyl  46-methoxy-5-(N′-(2-methylphenyl)- L D V OH urea)-2-pyridylacetyl  53-isoquinolinecarbonyl L E V OH  6 3-isoquinolinecarbonyl L hydroxyl- VOH aminocarb- onylmethyl  7 3-isoquinolinecarbonyl L S V OH  83-isoquinolinecarbonyl L (N-Bn)-H V OH  9 3-isoquinolinecarbonyl L C VOH  10 3-isoquinolinecarbonyl L tetrazol-5- V OH yl-methyl  113-isoquinolinecarbonyl L D — NH-CyM  12 3-isoquinolinecarbonyl L D — OH 13 3-(4-hydroxyphenyl)propionyl L D V OMe (R² = Me)  143-(4-hydroxyphenyl)propionyl L D — NH-CyM  153-(4-hydroxyphenyl)propionyl L d — NHi-Bu  163-(4-hydroxyphenyl)propionyl I d — NHi-Bu  173-(4-hydroxyphenyl)propionyl I D — NHi-Bu  183-(4-hydroxyphenyl)propionyl (N-Me)-L D V OMe  193-(4-hydroxyphenyl)propionyl L D V OMe  20 3-(4-hydroxyphenyl)propionylL D (N-Me)-V OMe  21 3-(4-hydroxyphenyl)propionyl L 1-carboxy- V OMeethyl  22 3-(4-hydroxyphenyl)propionyl L (N-Me)-D V OMe  23tetrahydro-3-isoquinolinecarbonyl L D — OH  24 3-phenylpropionyl L D —NH-CyM  25 4-phenylbutyryl L D — NH-CyM  26 5-phenylpentanoyl L D —NH-CyM  27 tetrahydro-3-isoquinolinecarbonyl L (N-Bn)-H V OH  28 acetyl(N-Bn)-L D V OMe  29 acetyl (N-phen- D V OMe ethyl)-L  303-phenylpropionyl (N-phen- D V OMe ethyl)-L  31tetrahydro-3-isoquinolinecarbonyl L E V OH  32 3-isoquinolinecarbonyl LD V/P OH  33 tetrahydro-3-isoquinolinecarbonyl L D V/P OH  34phenylacetyl L D V/P OH  35 phenylacetyl L D V/P OMe  363-phenylpropionyl L D V/P OH  37 3-phenylpropionyl L D V/P OMe  383-(4-hydroxyphenyl)propionyl L D V/P OH  39 3-(4-hydroxyphenyl)propionylL D V/P OMe  40 Boc L D V/P OMe  41 2-quinolinecarbonyl L D V/P OMe  42phenylacetyl L D V/pipeco- OH linyl  43 phenylacetyl L D V/n-butyl OH 44 2-quinolinecarbonyl L D V/n-butyl OH  45 4-methoxyphenylacetyl(N-Me)-L D V NHMe  46 3-(4-hydroxyphenyl)propionyl (N-Me)-L D V NHMe  47benzylaminocarbonyl L D V NHMe  48 p-tolylaminocarbonyl L D V NHMe  49phenylacetyl n-propyl D V NHMe  50 phenylacetyl L D V NHNaP  51phenylacetyl L D n-propyl NHMe  52 2-quinolinecarbonyl L D n-propyl NHMe 53 phenylacetyl L D 2-butyl NH₂  54 phenylacetyl L D 2-butyl OMe  55phenylacetyl L D 2-butyl NHMe  56 2-quinolinecarbonyl L D 2-butyl OMe 57 2-quinolinecarbonyl L D 2-butyl NHMe  58 1,2,3,4-tetrahydro-2- L D2-butyl NHMe quinolinecarbonyl  59 2-quinolinecarbonyl L D (O-Me)-T NHMe 60 2-quinolinecarbonyl L D T NHt-Bu  61 2-quinolinecarbonyl L D Tmorpho- lino  62 Boc L D T NHt-Bu  63 2-N-Boc-amino-1,2,3,4-tetrahydro-L D V OH 2-naphthoyl  64 3-phenylpropionyl L D V OH  653-(4-hydroxyphenyl)-2-bis- L D V OH (methylsulfonyl)aminopropionyl  663-(4-hydroxyphenyl)-2-N-Boc- L D V OH aminopropionyl  672-amino-1,2,3,4-tetrahydro-2- L D V OH naphthoyl TFA salt  68 Boc D V —OH  69 3-isoquinolinecarbonyl L D V OH  70 3-isoquinolinecarbonyl D V —OH  71 1,2,3,4-tetrahydro-3- D V — OH isoquinolinecarbonyl  72 naphthoylL D V OH  73 1,2,3,4-tetrahydro-2-naphthoyl L D V OH  74 naphthoyl D V —OH  75 1,2,3,4-tetrahydro-2-naphthoyl D V — OH  76 5-phenylpentanoyl D V— OH  77 2-pyridinecarbonyl L D V OH  78 2-pyridinecarbonyl D V — OH  793-tetrahydrofurancarbonyl L D V OH  80 2-tetrahydrofurancarbonyl L D VOH  81 3-isoquinolinecarbonyl F D V OH  82 3-isoquinolinecarbonyl A D VOH (R² = Me)  83 3-isoquinolinecarbonyl cyclohexyl D V OH  841,2,3,4-tetrahydro-3- cyclohexyl D V OH isoquinolinecarbonyl  853-isoquinolinecarbonyl cyclohexyl- D V OH methyl  861,2,3,4-tetrahydro-3- cyclohexyl- D V OH isoquinolinecarbonyl methyl  873-isoquinolinecarbonyl D F — OH  88 1,2,3,4-tetrahydro-3- D L — OHisoquinolinecarbonyl  89 3-isoquinolinecarbonyl D L — OH  901,2,3,4-tetrahydro-3- L D L OH isoquinolinecarbonyl  913-isoquinolinecarbonyl L D L OH  92 1,2,3,4-tetrahydro-3- L D F OHisoquinolinecarbonyl  93 3-isoquinolinecarbonyl L D F OH  942-quinolinecarbonyl L D V OH  95 3,3-diphenylpropionyl L D V OH  961,2,3,4-tetrahydro-3- A D V OH isoquinolinecarbonyl  973-isoquinolinecarbonyl A D V OH  98 5-phenylpentanoyl L D V OH  99indole-2-carbonyl L D V OH 100 3-(4-hydroxy)phenylpropionyl L D — NHi-Bu101 benzoyl L D — NHi-Bu 102 5-phenylpentanoyl L D — NHi-amyl 1033-(4-hydroxy)phenylpropionyl L D — NHi-amyl 104 6-phenylhexanoyl L D VOH 105 benzoyl L D V OH 106 5-phenylpentanoyl L D — NHi-Bu 107N-phenylsuccinamoyl L D V OH 108 N-4-fluorophenylsuccinamoyl L D V OH109 N-methyl-N-phenylsuccinamoyl L D V OH 110 1,2,3,4-tetrahydro-2- L D— NHi-amyl quinolinecarbonyl 111 N-phenylsuccinamoyl L D — NHi-Bu 1123-phenylpropyl (N-Me)-L (O-Me)-D V OMe 113 benzoyl (N-Me)-L D V OH 1141,2,3,4-tetrahydro-2- L D V NHHex quinolinecarbonyl 1151,2,3,4-tetrahydro-2- L D V 4-phenyl- quinolinecarbonyl piperidine 1163-(4-hydroxy)phenylpropionyl L D — NHHex 1173-(4-hydroxy)phenylpropionyl L D — N(iBu)₂ 1183-(4-hydroxy)phenylpropionyl L D — N(iBu)₂ 1193-(4-hydroxy)phenylpropionyl L D V NHHex 120 1,2,3,4-tetrahydro-2- L D VNMePh quinolinecarbonyl 121 2-quinolinecarbonyl L D V NMePh 1221,2,3,4-tetrahydro-2- L D V NH-4-fluoro- quinolinecarbonyl phenyl 1232-quinolinecarbonyl L D V NH-4-fluoro- phenyl 124 1,2,3,4-tetrahydro-2-L D V NHPh quinolinecarbonyl 125 2-quinolinecarbonyl L D V NHPh 1262-pyridinecarbonyl (N-Me)-L D V NHMe 127 2-quinolinecarbonyl L D V4-phenyl- piper- azinyl 128 4-methoxybenzoyl (N-Me)-L D V NHMe 129phenylacetyl Y D V NHMe 130 phenylacetyl P D V NHMe 131 phenylecetyl R DV NHMe 132 phenylacetyl N D V NHMe 133 2-N-Boc-amino-1,2,3,4- D V — NHMetetrahydro-2-naphthoyl 134 2-N-phenylacetylamino-1,2,3,4- D V — NHMetetrahydro-2-naphthoyl 135 Boc D P G OH 136 phenylacetyl D P G OH 137phenylacetyl L D — N-[bis- (carboxy)- methyl]- pipecolin- amido 138phenylacetyl L D P NH-[bis- (carboxy)- methyl] 139 phenylacetyl L D — N-[carboxy- methyl]- pipecolin- amide 140 3-phenylpropionyl (N-Me)-L D VOMe 141 4-hydroxyphenylacetyl (N-Me)-L D V OMe 142 2-quinolinecarbonyl(N-Me)-L D V OMe 143 4-phenylbutyryl (N-Me)-L D V OMe 1444-(N′-2-hydroxy- L D V/P OH phenylurea)phenylacetyl 145 PUPA L D V/P OH146 4-(N′-2-hydroxy- M D V/P OH phenylurea)phenylacetyl 1473-methoxy-4-N′- L D V/P NH₂ phenylurea)phenylacetyl 148 2-MPUPA L D V/PNH₂ 149 Boc D V P OH 150 5-phenylpentanoyl D V P OH 1512-allyl-4-phenylbutyryl V P — OH 152 acetyl F L D/V OH 153 benzoyl F LD/V OH 154 1,2,3,4-tetrahydro-3- L D V OMe isoquinolinecarbonyl 1554-phenylbutyryl L D V OH 156 3-isoquinolinecarbonyl L D V OMe 1573-isoquinolinecarbonyl L D — NHi-Bu 158 2-quinolinecarbonyl L D V Ot-Bu159 2-quinolinecarbonyl L (O-Bn)-D V OH 160 2-quinolinecarbonyl L D D OH161 4-phenylbutyryl L D — NHi-Bu 162 3-phenylpropionyl L D — NHi-B 163benzoyl G L D NHi-Bu 164 2-quinolinecarbonyl L D V NHMe 1654-methoxybenzoyl L D — NHi-Bu 166 4-phenylbutylyl L D V OMe 167 Boc L DV/M OMe 168 2-quinolinecarbonyl L D V/M OMe 169 N-n-butylaminocarbonyl DV — OMe 170 2-quinolinecarbonyl L D T OMe 171 N-t-butylaminocarbonyl L D— NHi-Bu 172 benzoyl G D V OMe 173 benzoyl G (O-Me)-D V OMe 1742-quinolinecarbonyl L D — NH(1-hy- droxy- methyl-2- methyl- propyl) 1752-quinolinecarbonyl L D V morpho- lino 176 4-methoxyphenylacetyl L D TOMe 177 4-methoxyphenylsulfonyl L D T OMe 178 2-quinolinecarbonyl L D VNH₂ 179 2-quinolinecarbonyl (N-Me)-L D V NHMe 180 phenylacetyl (N-Me)-LD V NHMe 181 phenylacetyl L D V NHMe 182 3-phenylpropionyl (N-Me)-L D VNHMe 183 phenylacetyl M D V NHMe 184 3-phenylpropionyl (N-Me)-L D V NHMe185 2-quinolinecarbonyl L D A NHMe (R² = Me) 186 2-quinolinecarbonyl L DV/M OH 187 phenylaminocarbonyl L D V NHMe 188 4-hydroxyphenylacetyl(N-Me)-L D V NHMe 189 phenylsulfonyl L D V NHMe 190 phenylacetyl L D(O-Me)-T OMe 191 phenylacetyl L D T OMe 192 phenylacetyl L D (O-Bn)-TOMe 193 phenylacetyl L D (O-Ac)-T OMe 194 phenylacetyl V D V NHMe 1952-quinolinecarbonyl L D T On-Bu 196 phenylacetyl L D V On-Bu 1972-quinolinecarbonyl L D T NH(4- methoxy- benzyl) 198 2-quinolinecarbonylL D — NH(3,3- dimethyl-n- butyl) 199 PUPA l D V/P NH₂ 200 PUPA L d V/PNH₂ 201 PUPA L D V/P NH₂ 202 2-MPUPA (N-6- D V/P OH amino- hexanoyl)-K203 PUPA L D V OH 204 PUPA L D V NHMe 205 PUPA L D V NHi-Bu 206 2-MPUPAL D V/P OH 207 2-MPUPA L D phenyl OH 208 PUPA L D V/P NH₂ 209 PUPA l DV/P NH₂ 210 PUPA L d V/P NH₂ 211 PUPA L D v/P NH₂ 212 PUPA l d v/p NH₂213 PUPA L D — NHBn 214 PUPA L D — morpho- lino 215 PUPA L D — NHi-Pr216 PUPA L D — NHCy 217 PUPA L D — NHi-Bu 218 PUPA L D — piperidinyl 2192-MPUPA M D D NH₂ 220 2-MPUPA M D L NH₂ 221 2-MPUPA M D V NH₂ 2222-MPUPA M D l NH₂ 223 2-MPUPA M D E NH₂ 224 2-MPUPA M D T NH₂ 2252-MPUPA M D M NH₂ 226 2-MPUPA M D n NH₂ 227 2-MPUPA M D e NH₂ 2282-MPUPA M D W NH₂ 229 2-MPUPA M D s NH₂ 230 2-MPUPA L D D NH₂ 2312-MPUPA L D L NH₂ 232 2-MPUPA L D V NH₂ 233 2-MPUPA L D l NH₂ 2342-MPUPA L D E NH₂ 235 2-MPUPA L D T NH₂ 236 2-MPUPA L D M NH₂ 2372-MPUPA L D n NH₂ 238 2-MPUPA L D e NH₂ 239 2-MPUPA L D W NH₂ 2402-MPUPA L D s NH₂ 241 2-MPUPA P D D NH₂ 242 2-MPUPA P D L NH₂ 2432-MPUPA P D V NH₂ 244 2-MPUPA P D l NH₂ 245 2-MPUPA P D E NH₂ 2462-MPUPA P D T NH₂ 247 2-MPUPA P D M NH₂ 248 2-MPUPA P D n NH₂ 2492-MPUPA P D e NH₂ 250 2-MPUPA P D W NH₂ 251 2-MPUPA P D s NH₂ 2522-MPUPA T D D NH₂ 253 2-MPUPA T D L NH₂ 254 2-MPUPA T D V NH₂ 2552-MPUPA T D l NH₂ 256 2-MPUPA T D E NH₂ 257 2-MPUPA T D T NH₂ 2582-MPUPA T D M NH₂ 259 2-MPUPA T D n NH₂ 260 2-MPUPA T D e NH₂ 2612-MPUPA T D W NH 262 2-MPUPA T D s NH₂ 263 2-MPUPA E D D NH₂ 264 2-MPUPAE D L NH₂ 265 2-MPUPA E D V NH₂ 266 2-MPUPA E D l NH₂ 267 2-MPUPA E D ENH₂ 268 2-MPUPA E D T NH₂ 269 2-MPUPA E D M NH₂ 270 2-MPUPA E D n NH₂271 2-MPUPA E D e NH₂ 272 2-MPUPA E D W NH₂ 273 2-MPUPA E D s NH₂ 2742-MPUPA C D V NH₂ 275 2-MPUPA S D D NH₂ 276 2-MPUPA S D L NH₂ 2772-MPUPA S D V NH₂ 278 2-MPUPA S D l NH₂ 279 2-MPUPA S D E NH₂ 2802-MPUPA S D T NH₂ 281 2-MPUPA S D M NH₂ 282 2-MPUPA S D n NH₂ 2832-MPUPA S D e NH₂ 284 2-MPUPA S D W NH₂ 285 2-MPUPA S D s NH₂ 2862-MPUPA l D D NH₂ 287 2-MPUPA l D L NH₂ 288 2-MPUPA l D V NH₂ 2892-MPUPA l D l NH₂ 290 2-MPUPA l D E NH₂ 291 2-MPUPA l D T NH₂ 2922-MPUPA l D M NH₂ 293 2-MPUPA l D n NH₂ 294 2-MPUPA l D e NH₂ 2952-MPUPA l D W NH₂ 296 2-MPUPA l D s NH₂ 297 2-MPUPA Q D D NH₂ 2982-MPUPA Q D L NH₂ 299 2-MPUPA Q D V NH₂ 300 2-MPUPA Q D l NH₂ 3012-MPUPA Q D E NH₂ 302 2-MPUPA Q D T NH₂ 303 2-MPUPA Q D M NH₂ 3042-MPUPA Q D n NH₂ 305 2-MPUPA Q D e NH₂ 306 2-MPUPA Q D W NH₂ 3072-MPUPA Q D s NH₂ 308 2-MPUPA M E D NH₂ 309 2-MPUPA M E V NH₂ 3102-MPUPA L E D NH₂ 311 2-MPUPA L E V NH₂ 312 2-MPUPA P E D NH₂ 3132-MPUPA P E V NH₂ 314 2-MPUPA T E D NH₂ 315 2-MPUPA M D V/P OH 3164-(N′-2-pyridylurea)phenylacetyl L D V/P OH 3173-methoxy-4-(N′-2-methylphenyl)- L D V/P NH₂ urea)phenylacetyl 318 PUPAL D V morpho- lino 319 PUPA L D V NHi-Pr 320 PUPA L D V NHCy 321 PUPA LD V NHBn 322 PUPA L D V piperidinyl 323 PUPA L D V NHi-Bu 324 PUPA L DV/P NHCy 325 PUPA L D V/P piperidinyl 326 PUPA L D V/P NHBn 327 PUPA L DV/P NHi-Pr 328 PUPA L D V/P NHi-Bu 329 2-MPUPA L D V morpho- lino 3303-(4-hydroxyphenyl) pipecolyl D — NHi-Bu 3313-(4-hydroxyphenyl)-propionyl P D — NHi-Bu 332 3-isoquinolinecarbonyl L(N-3- — OH methyl-2- butyroyl)- N 333 4-methylpentanoyl D — — NHCyM 334Cbz —CH₂CH₂— (N— V OMe (N of A²) CH₂CH₂— (C of A¹)-D 3353-(4-hydroxyphenyl)propionyl —CH₂CH₂— (N— V OMe (N of A²) CH₂CH₂— (C ofA¹)-D 336 4-(2-fluorophenyl- L D V/P OH urea)phenylacetyl 337 2-MPUPA LD V/P/S OH 338 2-MPUPA L D V/P/S/T OH 339 2-MPUPA V L P/D OH 340 2-MPUPAv I p/d OH 341 2-MPUPA L P V/D OH 342 2-MPUPA P D — OH 343 hydrogen p vd/I 2-MPUBA 344 hydrogen v d I 2-MPUBA 345 2-MPUPA L D I OH 3464-(N-(6-methyl-2- L D V/P OH pyridyl)urea)phenylacetyl 3474-(N-2-fluorophenyl- L D V/P OH urea)phenylacetyl 348 4-phenylbutyroyl(N-Me)-L D V NHMe 349 phenylacetyl S D V NHMe 350 phenylacetyl K D VNHMe 351 phenylacetyl L D A NHMe (R² = Me) 352 phenylacetyl L D (O-Bn)-SNHMe 353 2-quinolinecarbonyl L D (O-Bn)-S NHMe 354 Boc L D T NHBu 355Boc L D V/P OH 356 2-quinolinecarbonyl L D V/P OH 3574-(N′-2-pyridylurea)phenylacetyl L D V/P NH₂ 358 2-MPUPA L D*THAM V/POTHAM 359 2-MPUPA L D*Na V/P ONa 360 2-MPUPA L(l) Het¹ — — 361 2-MPUPA IHet¹ — — 362 2-MPUPA L(l) Het² — — 363 2-MPUPA L(l) Het³ — — 364 2-MPUPAL(l) Het⁴ — — 365 2-MPUPA L(l) Het⁵ — — 366 9-fluorenylmethoxycarbonyl LD V OH 367 3-methoxyphenylacetyl L D V OH 3683-(3-methylindolyl)propionyl L D V OH 369 2-phenyl-3-methyl-pyrazol-4- LD V OH ylcarbonyl 370 6-methylbenzpyrimidon-2- L D V OH ylcarbonyl 3714-oxo-4,5,6,7-tetrahydrobenzo- L D V OH [b]furan-3-ylcarbonyl 3723-(5-(phenylacetylenyl)pyri- L D V OH dinecarbonyl 3733-(2-phenylthio)-pyridinecarbonyl L D V OH 374 4-propylbenzoyl L D V OH375 4-(2-(3-pyridiyl))thiazolecarbonyl L D V OH 3764-(2-(4-pyridinyl)thiazolecarbonyl L D V OH 377 5-(2-(3-pyridinyl))thio-L D V OH phenesulfonyl 378 5-(2-(1-pyrrolyl))pyridinecarbonyl L D V OH379 N,N-(4-trifloromethylpyridin-2- L D V OH yl)methylhydraziocarbonyl380 2-quinoxalinylaminocarbonyl L D V OH 381N-(4-trifluoromethylpyridin-2- L D V OH yl)piperazinocarbonyl 382S-(2-(2-trifluoromethyl)- L D V OH phenylsulfonyl)-tetrahydrothiophenesulfonyl 383 1-(4-chlorophenylmethyl)pyr- L D V OHrolidin-2-on-4-yl-carbonyl 384 1-(2-furanylmethy)lpyr- L D V OHrolidin-2-on-4-ylcarbonyl 385 2(1-pyrrolyl)benzoyl L D V OH 3866-chlorochroman-3-ylcarbonyl L D V OH 387 2,3-dihydrobenzofuran- L D VOH 5-ylcarbonyl 388 4,6-dimethylpyrazolo[1,5-c] L D V OHtriazin-3-ylcarbonyl 389 3,4-benzocyclohexanoyl L D V OH 390norbonylacetyl L D V OH 391 1,2,3,4-tetrahyro-9- L D V OHacridinylcarbonyl 392 5,6,7,8-tetrahydronaphthyl- L D V OH aminocarbonyl393 3-(2-(4-methylthiophenoxy))- L D V OH pyridinecarbonyl 3942-(6-methoxynaphth-2-yl)propionyl L D V OH 395 (2-naphthyloxy)acetyl L DV OH 396 3-quinuclidinylaminocarbonyl L D V OH 397 2-(1,2,3,4-tetra- L DV OH hydroisoquinoline)carbonyl 398 adamantan-2-ylcarbonyl L D V OH 399(2-pyridyl)acetyl L D V OH 400 6-methylcyclohexen-2-ylcarbonyl L D V OH401 (3-quinolinyl)acetyl L D V OH 402 4-(2-butyl)phenylaminocarbonyl L DV OH 403 1,4-dihydro-1-ethyl-7-methyl-4- L D V OHoxo-1,8-naphthyrdin-3-ylcarbonyl 404 (2-thienyl)acetyl L D V OH 4054-(2-propyl)benzoyl L D V OH 406 3,4-methylenedoxybenzoyl L D V OH 4072-(5-(2-pyridyl))thiophenecarbonyl L D V OH 408 N-iminodibenzylcarbonylL D V OH 409 2-MPUPA P D l NHMe 410 2-MPUPA P D l OMe 411 2-MPUPA P D lOH 412 2-MPUPA —CH₂CH₂— D l OH (N of R¹) 413 2-MPUPA P E — NMe 4142-MPUPA P E l NMe 415 2-MPUPA P E l OH 416 2-MPUPA P E — OHwhere Het¹, Het², Het³, Het⁴, and Het⁵ in Table 1 are defined below:

The more preferred compounds of formula (I) are selected from the groupconsisting of compound numbers 1, 2, 4, 144, 145, 146, 147, 148, 206,315, 316, 317, 337, 338, 345, 346, 347, 357, 358 and 359 as identifiedin Table 1. Even more preferred compounds of formula (I) are selectedfrom the group consisting of compound numbers 1, 206, 316, 358 and 359as identified in Table 1. The most preferred compounds of formula (I)are selected from the group consisting of compound numbers 358 and 359as identified in Table 1.

Other compounds of this invention are compounds of formula II:K—(Y¹)—(Y²)—(Y³)_(n)—J  (II)and pharmaceutically acceptable derivatives thereof, wherein;

K is selected from the group consisting of hydrogen, alkyl, aliphaticacyl, aroyl, aralkylcarbonyl, heterocycloyl, sulfonyl, aralkylcarbonyl,heterocycloalkylcarbonyl, alkoxycarbonyl, aralkyloxycarbonyl,heterocycloalkoxycarbonyl, alkylaminocarbonyl and aralkylaminocarbonyl;

J is selected from the group consisting of alkoxy; aryloxy; aralkyloxy;hydroxyl; amino; alkylamino optionally substituted with hydroxy,aminocarbonyl, N-alkylaminocarbonyl, carboxy or alkoxycarbonyl;dialkylamino; cycloalkylamino; dicycloalkylamino; (alkyl)(aryl)amino;aralkylamino optionally substituted with carboxy; diaralkylamino;arylamino; and (mono- or bis-carboxylic acid)-substituted alkylamine;and

each Y, Y², Y³, R¹, A¹, A², A³, R², and n is independently as defined informula I above.

Compounds of this invention may be synthesized using any conventionaltechnique. Preferably, these compounds are chemically synthesized fromreadily available starting materials, such as α-amino acids and theirfunctional equivalents. Modular and convergent methods for the synthesisof these compounds are also preferred. In a convergent approach, forexample, large sections of the final product are brought together in thelast stages of the synthesis, rather than by incremental addition ofsmall pieces to a growing molecular chain.

According to one embodiment, compounds of the present invention may besynthesized in the following manner. A protected amino acid orfunctional equivalent is coupled to an appropriate activated estermoiety. The coupled product, if suitably functionalized, may be furtherreacted with yet another activated ester moiety. This material can befurther manipulated to give the desired compounds of the invention. Ateach step of the above sequence, the ester can be hydrolyzed to thecorresponding acid to give another compound of the invention. This acidmay also be converted to a corresponding acid derivative by standardmethods.

Alternatively, the activated ester moieties mentioned above can beattached together first, then the resulting compound can be attached toadditional amino acids or their functional group equivalents. At thispoint the final manipulations and/or necessary deprotection steps can beperformed.

In another embodiment, under suitable conditions the desiredfunctionalities can be incorporated (protected or unprotected) in one ofthe activated ester moieties. That ester is then coupled with an aminoacid derivative or a moiety consisting of an amino acid derivativepreviously coupled to an activated ester. The resulting product can thenbe subjected to any deprotection steps, If necessary, to give compoundsof the invention.

Alternatively, the compounds of this invention may be synthesized usingsolid support techniques. The core amino acid or their functionalequivalent groups are assembled using standard reiterative couplingmethodology on a resin. When the desired core is complete, the resultingfragment can be coupled with an activated ester moiety and/or thetethered end of the fragment may be further derivatized to give thedesired product. Appropiate protection/deprotection methods may be usedat any point during the synthetic sequence.

The compounds of this invention may also be modified by appendingappropriate functionalities to enhance selective biological properties.Such modifications are known in the art and include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion. Examples of these modificationsinclude, but are not limited to, esterification with polyethyleneglycols, derivatization with pivolates or fatty acid substituents,conversion to carbamates, hydroxylation of aromatic rings, andheteroatom-substitution in aromatic rings.

As used throughout this application, the term “patient” refers tomammals, including humans. And the term “cell” refers to mammaliancells, including human cells.

Once synthesized, the activities and VLA-4 specificities of thecompounds according to this invention may be determined using in vitroand in vivo assays.

For example, the cell adhesion inhibitory activity of these compoundsmay be measured by determining the concentration of inhibitor requiredto block the binding of VLA-4-expressing cells to fibronectin- orCS1-coated plates. In this assay microtiter wells are coated with eitherfibronectin (containing the CS-1 sequence) or CS-1. If CS-1 is used, itmust be conjugated to a carrier protein, such as bovine serum albumin,in order to bind to the wells. Once the wells are coated, varyingconcentrations of the test compound are then added together withappropriately labelled, VLA-4-expressing cells. Alternatively, the testcompound may be added first and allowed to incubate with the coatedwells prior to the addition of the cells. The cells are allowed toincubate in the wells for at least 30 minutes. Following incubation, thewells are emptied and washed. Inhibition of binding is measured byquantitating the fluorescence or radioactivity bound to the plate foreach of the various concentrations of test compound, as well as forcontrols containing no test compound.

VLA-4-expressing cells that may be utilized in this assay include Ramoscells, Jurkat cells, A375 melanoma cells, as well as human peripheralblood lymophocytes (PBLs). The cells used in this assay may befluorescently or radioactively labelled.

A direct binding assay may also be employed to quantitate the inhibitoryactivity of the compounds of this invention. In this assay, a VCAM-IgGfusion protein containing the first two immunoglobulin domains of VCAM(D1D2) attached above the hinge region of an IgG1 molecule (“VCAM2D-IgG”), is conjugated to a marker enzyme, such as alkaline phosphatase(“AP”). The synthesis of this VCAM-IgG fusion is described in PCTpublication WO 90/13300, the disclosure of which is herein incorporatedby reference. The conjugation of that fusion to a marker enzyme isachieved by cross-linking methods well-known in the art.

The VCAM-IgG enzyme conjugate is then placed in the wells of amulti-well filtration plate, such as that contained in the MilliporeMultiscreen Assay System (Millipore Corp., Bedford, Mass.). Varyingconcentrations of the test inhibitory compound are then added to thewells followed by addition of VLA-4-expressing cells. The cells,compound and VCAM-IgG enzyme conjugate are mixed together and allowed toincubate at room temperature.

Following incubation, the wells are vacuum drained, leaving behind thecells and any bound VCAM. Quantitation of bound VCAM is determined byadding an appropriate calorimetric substrate for the enzyme conjugatedto VCAM-IgG and determining the amount of reaction product. Decreasedreaction product indicates increased binding inhibitory activity.

In order to assess the VLA-4 inhibitory specificity of the compounds ofthis invention, assays for other major groups of integrins, i.e., β2 andβ3, as well as other β1 integrins, such as VLA-5, VLA-6 and α4β7 areperformed. These assays may be similar to the adhesion inhibition anddirect binding assays described above, substituting the appropriateintegrin-expressing cell and corresponding ligand. For example,polymorphonuclear cells (PMNs) express β2 integrins on their surface andbind to ICAM. β3 integrins are involved in platelet aggregation andinhibition may be measured in a standard platelet aggregation assay.VLA-5 binds specifically to Arg-Gly-Asp sequences, while VLA-6 binds tolaminin. α4β7 is a recently discovered homologue of VLA-4, which alsobinds fibronectin and VCAM. Specificity with respect to α4β7 isdetermined in a binding assay that utilizes the above-describedVCAM-IgG-enzyme marker conjugate and a cell line that expresses α4β7,but not VLA-4, such as RPMI-8866 cells.

Once VLA-4-specific inhibitors are identified, they may be furthercharacterized in vivo assays. One such assay tests the inhibition ofcontact hypersensitivity in an animal, such as described by P. L.Chisholm et al., “Monoclonal Antibodies to the Integrin α-4 SubunitInhibit the Murine Contact Hypersensitivity Response”, Eur. J. Immunol.,23, pp. 682-688 (1993) and in “Current Protocols in Immunology”, J. E.Coligan, et al., Eds., John Wiley & Sons, New York, 1, pp. 4.2.1-4.2.5(1991), the disclosures of which is herein incorporated by reference. Inthis assay, the skin of the animal is sensitized by exposure to anirritant, such as dinitrofluorobenzene, followed by light physicalirritation, such as scratching the skin lightly with a sharp edge.Following a recovery period, the animals are re-sensitized following thesame procedure. Several days after sensitization, one ear of the animalis exposed to the chemical irritant, while the other ear is treated witha non-irritant control solution. Shortly after treating the ears, theanimals are given various doses of the VLA-4 inhibitor by subcutaneousinjection. In vivo inhibition of cell adhesion-associated inflammationis assessed by measuring the ear swelling response of the animal in thetreated versus untreated ear. Swelling is measured using calipers orother suitable instrument to measure ear thickness. In this manner, onemay identify those inhibitors of this invention which are best suitedfor inhibiting inflammation.

Another in vivo assay that may be employed to test the inhibitors ofthis invention is the sheep asthma assay. This assay is performedessentially as described in W. M. Abraham et al., “α-Integrins MediateAntigen-induced Late Bronchial Responses and Prolonged AirwayHyperresponsiveness in Sheep”, J. Clin. Invest., 93, pp. 776-87 (1994),the disclosure of which is herein incorporated by reference. This assaymeasures inhibition of Ascaris antigen-induced late phase airwayresponses and airway hyperresponsiveness in asthmatic sheep.

The compounds of the present invention may be used in the form ofpharmaceutically acceptable salts derived from inorganic or organicacids and bases. Included among such acid salts are the following:acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate,bisulfate, butyrate, citrate, camphorate, camphorsulfonate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate,hexanoate, hydrochloride, hydrobromide, hydroiodide,2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate,2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate,persulfate, 3-phenylpropionate, picrate, pivalate, propionate,succinate, tartrate, thiocyanate, tosylate and undecanoate. Base saltsinclude ammonium salts, alkali metal salts, such as sodium and potassiumsalts, alkaline earth metal salts, such as calcium and magnesium salts,salts with organic bases, such as dicyclohexylamine salts,N-methyl-D-glucamine, tris(hydroxymethyl)methylamine and salts withamino acids such as arginine, lysine, and so forth. Also, the basicnitrogen-containing groups can be quaternized with such agents as loweralkyl halides, such as methyl, ethyl, propyl, and butyl chloride,bromides and iodides; dialkyl sulfates, such as dimethyl, diethyl,dibutyl and diamyl sulfates, long chain halides such as decyl, lauryl,myristyl and stearyl chlorides, bromides and iodides, aralkyl halides,such as benzyl and phenethyl bromides and others. Water or oil-solubleor dispersible products are thereby obtained.

The compounds of the present invention may be formulated intopharmaceutical compositions that may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques.

The pharmaceutical compositions of this invention comprise any of thecompounds of the present invention, or pharmaceutically acceptablederivatives thereof, together with any pharmaceutically acceptablecarrier. The term “carrier” as used herein includes acceptable adjuvantsand vehicles. Pharmaceutically acceptable carriers that may be used inthe pharmaceutical compositions of this invention include, but are notlimited to, ion exchangers, alumina, aluminum stearate, lecithin, serumproteins, such as human serum albumin, buffer substances such asphosphates, glycine, sorbic acid, potassium sorbate, partial glyceridemixtures of saturated vegetable fatty acids, water, salts orelectrolytes, such as protamine sulfate, disodium hydrogen phosphate,potassium hydrogen phosphate, sodium chloride, zinc salts, colloidalsilica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-basedsubstances, polyethylene glycol, sodium carboxymethylcellulose,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers,polyethylene glycol and wool fat.

According to this invention, the pharmaceutical compositions may be inthe form of a sterile injectable preparation, for example a sterileinjectable aqueous or oleaginous suspension. This suspension may beformulated according to techniques known in the art using suitabledispersing or wetting agents and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are water, Ringer's solutionand isotonic sodium chloride solution. In addition, sterile, fixed oilsare conventionally employed as a solvent or suspending medium. For thispurpose, any bland fixed oil may be employed including synthetic mono-or di-glycerides. Fatty acids, such as oleic acid and its glyceridederivatives are useful in the preparation of injectables, as do naturalpharmaceutically-acceptable oils, such as olive oil or castor oil,especially in their polyoxyethylated versions. These oil solutions orsuspensions may also contain a long-chain alcohol diluent or dispersant,such as Ph. Helv or similar alcohol.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers which are commonly used includelactose and corn starch. Lubricating agents, such as magnesium stearate,are also typically added. For oral administration in a capsule form,useful diluents include lactose and dried corn starch. When aqueoussuspensions are required for oral use, the active ingredient is combinedwith emulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Alternatively, the pharmaceutical compositions of this invention may beadministered in the form of suppositories for rectal administration.These can be prepared by mixing the agent with a suitable non-irritatingexcipient which is solid at room temperature but liquid at the rectaltemperature and therefore will melt in the rectum to release the drug.Such materials include cocoa butter, beeswax and polyethylene glycols.

The pharmaceutical compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, the pharmaceutical compositions may beformulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutical compositions can be formulatedin a suitable lotion or cream containing the active components suspendedor dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated asmicronized suspensions in isotonic, pH adjusted sterile saline, or,preferably, as solutions in isotonic, pH adjusted sterile saline, eitherwith our without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutical compositions maybe formulated in an ointment such as petrolatum.

The pharmaceutical compositions of this invention may also beadministered by nasal aerosol or inhalation through the use of anebulizer, a dry powder inhaler or a metered dose inhaler. Suchcompositions are prepared according to techniques well-known in the artof pharmaceutical formulation and may be prepared as solutions insaline, employing benzyl alcohol or other suitable preservatives,absorption promoters to enhance bioavailability, fluorocarbons, and/orother conventional solubilizing or dispersing agents.

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated, and the particular mode of administration. It should beunderstood, however, that a specific dosage and treatment regimen forany particular patient will depend upon a variety of factors, includingthe activity of the specific compound employed, the age, body weight,general health, sex, diet, time of administration, rate of excretion,drug combination, and the judgment of the treating physician and theseverity of the particular disease being treated. The amount of activeingredient may also depend upon the therapeutic or prophylactic agent,if any, with which the ingredient is co-administered.

The dosage and dose rate of the compounds of this invention effective toprevent, suppress or inhibit cell adhesion will depend on a variety offactors, such as the nature of the inhibitor, the size of the patient,the goal of the treatment, the nature of the pathology to be treated,the specific pharmaceutical composition used, and the judgment of thetreating physician. Dosage levels of between about 0.001 and about 100mg/kg body weight per day, preferably between about 0.1 and about 10mg/kg body weight per day of the active ingredient compound are useful.

According to another embodiment compositions containing a compound ofthis invention may also comprise an additional agent selected from thegroup consisting of corticosteroids, bronchodilators, antiasthmatics(mast cell stabilizers), antiinflammatories, antirheumatics,immunosuppressants, antimetabolites, immunonodulators, antipsoriaticsand antidiabetics. Specific compounds within each of these classes maybe selected from any of those listed under the appropriate groupheadings in “Comprehensive Medicinal Chemistry”, Pergamon Press, Oxford,England, pp. 970-986 (1990), the disclosure of which is hereinincorporated by reference. Also included within this group are compoundssuch as theophylline, sulfasalazine and aminosalicylates(antiinflammatories); cyclosporin, FK-506, and rapamycin(immunosuppressants); cyclophosphamide and methotrexate(antimetabolites); and interferons (immunomodulators).

According to other embodiments, the invention provides methods forpreventing, inhibiting or suppressing cell adhesion-associatedinflammation and cell adhesion-associated immune or autoimmuneresponses. VLA4-associated cell adhesion plays a central role in avariety of inflammation, immune and autoimmune diseases. Thus,inhibition of cell adhesion by the compounds of this invention may beutilized in methods of treating or preventing inflammatory, immune andautoimmune diseases. Preferably the diseases to be treated with themethods of this invention are selected from asthma, arthritis,psoriasis, transplantation rejection, multiple sclerosis, diabetes andinflammatory bowel disease.

These methods may employ the compounds of this invention in amonotherapy or in combination with an anti-inflammatory orimmunosuppressive agent. Such combination therapies includeadministration of the agents in a single dosage form or in multipledosage forms administered at the same time or at different times.

In order that this invention may be more fully understood, the followingexamples are set forth. These examples are for the purpose ofillustration only and are not to be construed as limiting the scope ofthe invention in any way.

EXAMPLES

General Procedures for Amide Bond Formation in Solution:

Procedure A: Coupling with EDC/HOBt

A solution of carboxylic acid (1.2 eq.) in DMF at 0° C. was treated withHOBT (1.8 eq.) and EDC (1.4 eq.). The mixture was stirred at 0° C. for 1to 2 h and then the free amine (1.0 eq., neutralized with TEA or DIPEA)was added. After stirring at RT for more than 3 h, the reaction mixturewas diluted with ethyl acetate, washed with water (1×), 5% aqueouscitric acid (2×), sat. NaHCO₃ (2×), and brine (1×), dried (Na₂SO₄ orMgSO₄), and concentrated in vacuo.

Procedure B: Coupling using Activated Ester (N-hydroxysuccinate orChloride)

A solution of free amine (1-1.2 eq., neutralized with TEA or DIPEA) inCH₂Cl₂ was treated with activated ester or acyl halide (1 eq.) at 0° C.or RT. After stirring at RT for over 1 h, the reaction mixture waswashed with 5% aqueous citric acid (2×), sat. NaHCO₃ (2×), and brine(1×), dried (Na₂SO₄ or MgSO₄), and concentrated in vacuo.

General Procedure for Urea Formation in Solution:

Procedure C: Formation of Urea with Isocyanate and Amine.

A solution of amine (1 eq.) and TEA (1 eq.) in CH₂Cl₂ was treated withan isocyanate (1 eq.) and was stirred at RT for over 0.5 h. Afterconcentration in vacuo, the product was either used as is or purified bychromatography.

General Procedures for Deprotection in Solution:

Procedure D: Removal of BOC with TFA

A solution of tBuOC(O)NH—R (where R is alkyl optionally substituted withany number of suitable functional groups) in CH₂Cl₂ at 0° C. was treatedwith trifluoroacetic acid. The reaction was allowed to warm to RT andstirred for 1 to 2 h. After concentration in vacuo the resultingamine/TFA salt was stored and neutralized with TEA or DIPEA prior touse.

Procedure E: Removal of BOC with HCl

A solution of tBuOC(O)NH—R (where R is alkyl optionally substituted withany number of suitable functional groups) in dioxane at 0° C. wastreated with 4N HCl in dioxane. The reaction was allowed to warm to RTand stirred for 1 to 2 h. After concentration in vacuo the resultingamine/HCl salt was stored and neutralized with TEA or DIPEA prior touse.

Procedure F: Hydrogenation

A mixture of starting material and 10% Pd/C in methanol, water, ethylacetate, and/or DMF was vigorously stirred under hydrogen (40 to 50 psi)for more than 2 h at RT. The resulting mixture was filtered through aplug of Celite and the filtrate concentrated in vacuo.

General Procedures for Amide Bond Formation on Solid Support:

Procedure G: Coupling with DCC/HOBt

A mixture of resin (see below for preparation of resin MCB1),tBuOC(O)NH—AA_(x)—CO₂H (where AA is an amino acid or functionalequivalent) or R₂—CO₂H (10 eq.), HOBt (10 eq.), DCC (10 eq) andN-methylmorpholine (3 eq) in NMP was shaken for over 0.5 h at RT. Theresin was then washed with NMP (2×) and CH₂Cl₂ (3×).

Procedure H: Displacement from Resin with Amine

A mixture of resin and amine (xs) in DMF was shaken for 6 h at RT. Theresin was then washed with methanol (3×) and the combined solutionsconcentrated in vacuo.

General Procedures for Deprotection on Solid Support:

Procedure I: Removal of BOC with TFA/CH₂Cl₂

A mixture of resin and 50% TFA/CH₂Cl₂ was shaken for over 0.5 h at RT.The resin was then washed with CH₂Cl₂ (2×), isopropanol (1×), and CH₂Cl₂(3×).

Procedure J: HF with Scavengers

The protected product was treated with HF at −10 to 0° C. for over 1.5 hin the presence of anisole or thioanisole as scavenger. The HF wasremoved with a stream of nitrogen gas at 0° C.

Example 1 Synthesis of Common Intermediates

Succinimidyl 3-Isoquinolinecarboxylate (iQn-OSu):

A solution of 3-isoquinolinecarboxylic acid (1.2 eq) in DMF at 0° C. wastreated with EDC (1.4 eq.). The mixture was stirred at 0° C. for 1 to 2h and then N-hydroxysuccinimide (1.0 eq.) was added. After stirring atRT for more than 3 h, the reaction mixture was poured into 60% sat.NaHCO₃ and the product filtered: ¹H NMR (CDCl₃, 300 MHz, ppm) 9.35 (s,1H), 8.67 (s, 1H), 8.09 (m, 1H), 7.96 (m, 1H), 7.82 (m, 2H), 2.94 (s,4H).

Succinimidyl 2-Quinolinecarboxylate (Qn-OSu):

A solution of 2-quinoline carboxylic acid (1.2 eq.) in DMF at 0° C. wastreated with EDC (1.4 eq.). The mixture was stirred at 0° C. for 1 to 2h and then N-hydroxysuccinimide (1.0 eq.) was added. After stirring atRT for more than 3 h, the reaction mixture was poured into 60% sat.NaHCO₃ and the product filtered: ¹H NMR (CDCl₃, 300 MHz, ppm) 8.35 (d,1H), 8.27 (d, 1H), 8.19 (d, 1H), 7.87 (d, 1H), 7.80 (m, 1H), 7.68 (m,1H), 2.91 (s, 4H).

Methyl 4-Isocyanatophenylacetate (KCl):

A well-stirred cold solution of methyl p-aminophenylacetate (9.8 g, 59.4mmol) in CH₂Cl₂ (200 mL) and TEA (25 mL, 18 g, 178.2 mmol) was treatedwith COCl₂ (96 mL of 1.9 M solution in toluene) over 1 h. The reactionmixture was stirred at 0° C. for an additional 1 h. The reaction mixturewas concentrated and 3:1 ether/pet ether (125 mL) was added. The mixturewas filtered and the filtrate concentrated to give KCl as a brownliquid. The crude product was purified by distillation (118-120° C./1.0mm) to afford pure KCl (8.5 g, 75%) as a colorless liquid: ¹H NMR(CDCl₃, 300 MHz, ppm) 7.20 (d, J=8.4 Hz), 7.02 (d, J=8.4 Hz), 3.69 (s,3H), 3.48 (s, 2H).

4-Phenylureidophenylacetic acid:

4-Phenylureidophenylacetic acid was prepared using procedure C with4-amino-phenylacetic acid and phenyl isocyanate: ¹H NMR (CD₃SOCD₃, 300MHz, ppm) 8.72-8.64 (m, 2H), 7.44 (d, 2H), 7.36 (d, 2H), 7.28 (d, 2H),7.16 (d, 2H), 6.96 (t, 1H), 3.52 (s, 2H); m/z 272.

4-o-Tolylureidophenylacetic acid:

4-o-Tolylureidophenylacetic acid was prepared using procedure C with4-amino-phenylacetic acid and o-tolyl isocyanate: ¹H NMR (CD₃SOCD₃, 300MHz, ppm) 8.97 (s, 1H), 7.88 (s, 1H), 7.83 (d, 1H), 7.38 (d, 2H),7.17-7.09 (m, 4H), 6.92 (t, 1H), 3.48 (s, 2H), 2.23 (s, 3H); m/z 285.

4-(2-Fluorophenyl)ureidophenylacetic acid:

4-(2-Fluorophenyl)ureidophenylacetic acid was prepared using procedure Cwith 2-fluoroaniline and KCl: ¹H NMR (CD₃SOCD₃, 300 MHz, ppm) 9.00 (s,1H), 8.51 (d, 2.4 Hz, 1H), 8.14 (dd, 8.3 Hz, 1.5 Hz, 1H), 7.37 (d, 8.5Hz, 2H), 7.07-7.25 (m, 4H), 6.99 (m, 1H), 3.48 (s, 2H).

4-(2-Hydroxyphenylureido)phenylacetic acid:

4-(2-Hydroxyphenylureido)phenylacetic acid was prepared using procedureC with 2-hydroxyaniline and KCl: ¹H NMR (CD₃SOCD₃, 300 MHz, ppm) 9.90(s, 1H), 9.25 (s, 1H), 8.12 (s, 1H), 8.02 (bd, 1H), 7.37 (d, 2H), 7.13(d, 2H), 6.70-6.97 (m, 3H), 3.48 (s, 2H).

N-Succinimidyl 4-(2-(3-methylpyridylureido)phenylacetate:

Prepared in three steps as follows:

Procedure C with 2-amino-3-methylpyridine and KCl to give methyl4-(2-(3-methylpyridylureido)phenylacetate.

A solution of methyl 4-(2-(3-methylpyridylureido)phehylacetate (1 eq.)in methanol was treated with 1 N NaOH (2 eq.). The reaction was stirredfor 16 h, then acidified carefully with 1 N HCl to pH 7 then with aceticacid to pH 3. The product was filtered and washed with methanol thenether to give 4-(2-(3-methylpyridylureido)phenylacetic acid: ¹H NMR(CD₃SOCD₃, 300 MHz, ppm) 11.97 (s, 1H), 8.64 (brs, 1H), 8.31 (s, 1H),7.69 (m, 1H), 7.62 (d, 8.4 Hz, 2H), 7.33 (d, 8.4 Hz, 2H), 7.09 (m, 1H),3.62 (s, 2H), 2.38 (s, 3H); m/z 286.

A solution of 4-(2-(3-methylpyridylureido)phenylacetic acid (1 eq.),N-hydroxysuccinimide (1.2 eq.) and EDC (1.2 eq.) in DMF was made basic(pH 10) with TEA. After stirring at RT for over 12 h, the reaction waspoured into 60% sat NaHCO₃ and the product filtered: ¹H NMR (CD₃SOCD₃,300 MHz, ppm) 12.04 (s, 1H), 8.84 (s, 1H), 8.31 (s, 1H), 7.72 (m, 3H),7.42 (m, 2H), 7.10 (m, 1H), 4.18 (s, 2H), 2.98 (3, 4H), 2.38 (s, 3H);m/z 383.

N-Succinimidyl 4-(2-pyridylureido)phenylacetate:

Prepared in three steps as follows:

Procedure-C with 2-aminopyridine and KCl to give methyl4-(2-pyridylureido)phenylacetate: ¹H NMR (CDCl₃, 300 MHz, ppm) 8.20 (s,2H), 7.62-7.51 (m, 3H), 7.33 (d, 2H), 7.01 (d, 2H), 6.89-6.85 (m, 1H),3.70 (s, 3H), 3.59 (s, 2H).

A solution of methyl 4-(2-pyridylureido)phenylacetate (5.7 g, 20.0 mmol)in methanol (20 mL) was treated with 1 N NaOH (40 mL). The reaction wasstirred for 16 h, then acidified carefully with 1 N HCl to pH 7 thenwith acetic acid to pH 3. The product was filtered and washed withmethanol then ether to give 4-(2-pyridyl)ureidophenylacetic acid (4.7 g,87%) as a white powder: ¹H NMR (CD₃SOCD₃, 300 MHz, ppm) 10.62 (bs, 1H),9.53 (bs, 1H), 8.39 (d, 1H), 7.82 (t, 1H), 7.63-7.55 (m, 1H), 7.33-7.27(d, 2H), 7.14-7.08 (m, 1H), 3.62 (s, 3H).

A solution of 4-(2-pyridyl)ureidophenylacetic acid (1 eq.),N-hydroxysuccinimide (1.2 eq.) and EDC (1.2 eq.) in DMF was made basic(pH 10) with TEA. After stirring at RT for over 12 h, the reaction waspoured into 60% sat. NaHCO₃ and the product filtered: ¹H NMR (CD₃SOCD₃,300 MHz, ppm) 10.08 (s, 1H), 9.57 (s, 1H), 8.39 (m, 1H), 7.86 (m, 1H),7.62 (m, 3H), 7.38 (d, 2H), 7.12 (m, 1H), 4.15 (s, 2H), 2.91 (s, 4H);m/z 369.

3-Methoxy-4-phenylureidophenylacetic acid:

Prepared in six steps from 3-methoxy-4-nitrobenzoic acid as follows:

A mixture of 3-methoxy-4-nitrobenzoic acid (2.01 g, 10.2 mmol) andthionyl chloride (2.3 mL, 31.5 mmol) was stirred at 80-90° C. for 1.5 h.The reaction was concentrated and the residue diluted with ether. Theorganic solution was washed with sat. aq. NaHCO₃ (2×), H₂O, then sat.aq. NaCl, dried (MgSO₄) and concentrated to afford3-methoxy-4-nitrobenzoyl chloride (1.92 g, 87%) as a white solid: ¹H NMR(CDCl3, 300 MHz, ppm) 7.95-7.70 (m, 3H), 4.06 (s, 3H).

A cold (0° C.) solution of TMSCHN₂ (2 M in hexane, 1.5 mL, 3.0 mmol) andtriethylamine (420 μL, 3.0 mmol) was treated with a solution of3-methoxy-4-nitrobenzoyl chloride (0.52 g, 2.4 mmol) in acetonitrile(8.5 mL). The reaction was stirred at 0° C. for 24 h and thenconcentrated. The residue was slurried with sat. aq. NaHCO₃ and themixture extracted with ether (3×). The combined ether washes were washedwith water, then sat. aq. NaCl, dried (MgSO₄) and concentrated to affordω-diazo-3-methoxy-4-nitroacetophenone (0.53 g, 100%) as a yellow foam:¹H NMR (CDCl₃, 300 MHz, ppm) 7.88 (d, 10 Hz, 1H), 7.61 (s, 1H), 7.27 (d,10 Hz, 1H), 5.97 (s, 1H), 4.02 (s, 3H).

A refluxing solution of ω-diazo-3-methoxy-4-nitroacetophenone (7.95 g,35.9 mmol) in t-BuOH (100 mL) was treated with a filtered solution ofsilver benzoate (2.50 g, 10.9 mmol) in triethylamine (15 mL) dropwiseover 1 h. After refluxing for 45 min, decolorizing carbon was added andthe hot mixture filtered through a pad of Celite. The filtrate wasconcentrated and the residue diluted with ethyl acetate. The organicsolution was washed with 5% aq. NaHCO₃ (2×), H₂O, 5% aq. citric acid,H₂O, then sat. aq. NaCl, dried (MgSO₄) and concentrated to affordt-butyl 3-methoxy-4-nitrophenylacetate (8.92 g, 93%) as a brown oil: ¹HNMR (CDCl₃, 300 MHz, ppm) 7.83 (d, 8.3 Hz, 1H), 7.03 (s, 1H), 6.93 (d,8.3 Hz, 1H), 3.97 (s, 3H), 3.58 (s, 2H), 1.45 (s, 9H).

A mixture of t-butyl 3-methoxy-4-nitrophenylacetate (0.144 g, 0.539mmol) and 10% Pd on carbon (0.155 g) in ethyl acetate (8 mL) andmethanol (2 mL) was stirred under H₂ (40-60 psi) for 2 h. The mixturewas filtered through Celite and the filtrate concentrated to affordt-butyl 4-amino-3-methoxyphenylacetate (0.123 g, 96%) as a light yellowoil: ¹H NMR (CDCl₃, 300 MHz, ppm) 6.70 (m, 3H), 4.04 (bs, 2H), 3.84 (s,3H), 3.42 (s, 2H), 1.43 (s, 9H).

Procedure C with t-butyl 4-amino-3-methoxyphenylacetate and phenylisocyanate gave t-butyl 3-methoxy-4-phenylureidophenylacetate: ¹H NMR(CDCl₃, 300 MHz, ppm) 8.00 (d, 11 Hz, 1H) 7.65-6.94 (m, 7H), 6.80 (d,9.0 Hz, 1H), 6.74 (s, 1H), 3.68 (s, 3H), 3.45 (s, 2H), 1.44 (s, 9H).

A solution of t-butyl 3-methoxy-4-phenylureidophenylacetate (0.108 g,0.303 mmol) in trifluoroacetic acid (5.0 mL) was stirred for 30 min. Thereaction was concentrated and the residue coevaporated with methylenechloride (2×) then ether to afford 3-methoxy-4-phenylureidophenylaceticacid (0.090 g, 99%) as a white foam: ¹H NMR (CD₃SOCD₃, 300 MHz, ppm)9.28 (s, 1H), 8.18 (s, 1H), 8.02 (d, 7.5 Hz, 1H), 7.58-7.15 (m, 5H),6.91 (bm, 2H), 6.77 (d, 7.5 Hz, 1H), 3.85 (s, 3H), 3.49 (s, 2H).

N-Succinimidyl 3-methoxy-4-phenylureidophenylacetate:

A solution of 3-methoxy-4-phenylureidophenylacetic acid (1 eq.) in DMFat 0° C. was treated with EDC (1.1 eq.). The mixture was stirred at 0°C. for 1 to 2 h and then N-hydroxysuccinimide (1.1 eq.) was added. Afterstirring at RT for more than 3 h, the reaction mixture was poured into60% sat. NaHCO₃ and the N-succinimidyl3-methoxy-4-phenylureidophenylacetate filtered.

N-Succinimidyl 6-(2-methoxy-3-o-tolylureido)pyridylacetate:

Prepared in six steps from 2,6-dichloro-3-nitropyridine as follows:

A slurry of 2,6-dichloro-3-nitropyridine (92%, 9.9 g, 47 mmol) and K₂CO₃powder (6.5 g, 47 mmol) in methanol (100 mL) was stirred for a week atRT. The reaction was filtered and concentrated. The residue waspartitioned in ethyl acetate and 60% sat. aq. NaHCO₃. The organicsolution was washed with 60% sat. aq. NaHCO₃ (2×), H₂O, then sat. aq.NaCl, dried (MgSO₄) and concentrated to afford2-chloro-6-methoxy-5-nitropyridine and2-chloro-6-methoxy-3-nitropyridine (8.9 g, 100%) as a light yellowsolid: ¹H NMR (CDCl₃, 300 MHz, ppm) 8.31 (d, 8.3 Hz, 1H), 8.28 (d, 8.9Hz, 1H), 7.10 (d, 8.3 Hz, 1H), 6.82 (d, 8.9 Hz, 1H), 4.15 (s, 3H), 4.06(s, 3H).

A mixture of 2-chloro-6-methoxy-5-nitropyridine and2-chloro-6-methoxy-3-nitropyridine (8.9 g, 47 mmol), t-butyl methylmalonate (10 mL, 60 mmol), and NaH (95%, 3.1 g, 120 mmol) in THF (250mL) was stirred at RT for 24 h. The reaction was concentrated and theresidue treated with trifluoroacetic acid (200 mL) for 2 h. The reactionwas concentrated and the product separated by flash chromatography(silica gel, 95:5 hexane-ethyl acetate) to afford methyl6-(2-methoxy-3-nitro)pyridylacetate (3.3 g, 62%) as a yellow oil: ¹H NMR(CDCl₃, 300 MHz, ppm) 8.27 (d, 8.0 Hz, 1H), 7.04 (d, 8.0 Hz, 1H), 4.09(s, 3H), 3.85 (s, 2H), 3.75 (s, 3H).

A mixture of methyl 6-(2-methoxy-3-nitro)pyridylacetate (0.047 g, 0.21mmol) and 10% Pd on carbon (0.063 g) in ethyl acetate (2 mL) and ethanol(1 mL) was stirred under H₂ (40-50 psi) for 6 h. The mixture wasfiltered through Celite and the filtrate concentrated to afford methyl6-(2-methoxy-3amino)pyridylacetate (0.041 g, 100%) as a light yellowoil: ¹H NMR (CDCl₃, 300 MHz, ppm) 6.82 (d, 7.6 Hz, 1H), 6.65 (d, 7.6 Hz,1H), 3.94 (s, 3H), 3.70 (s, 3H), 3.65 (s, 2H).

Procedure C with methyl 6-(2-methoxy-3-amino)pyridylacetate and o-tolylisocyanate to give methyl 6-(2-methoxy-3-o-tolylureido)pyridylacetate:¹H NMR (CDCl₃, 300 MHz, ppm) 8.33 (d, 7.9 Hz, 1H), 7.51 (d, 7.8 Hz, 1H),7.41 (s, 1H), 7.17 (m, 2H), 7.08 (m, 2H), 6.77 (d, 7.9 Hz, 1H), 3.81 (s,3H), 3.71 (s, 3H), 3.67 (s, 2H), 2.20 (s, 3H).

A solution of methyl 6-(2-methoxy-3-o-tolylureido)pyridylacetate (0.023g, 0.070 mmol) in methanol (1.0 mL) was treated with 2 M LiOH (90 μL,0.18 mmol). The reaction was stirred for 18 h, diluted with H₂O (5.0 mL)and washed with ether (2×). The aqueous solution was then acidified with5% aq. citric acid. The product was filtered and washed with H₂O thenether to give 6-(2-methoxy-3-o-tolylureido)pyridylacetic acid (0.014 g,64%) as a white solid: ¹H NMR (CD₃OD, 300 MHz, ppm) 8.50-8.25 (m, 3H),7.60 (bd, 1H), 7.28-7.00 (m, 3H), 4.01 (s, 3H), 3.69 (s, 2H), 2.30 (s,3H); MS, m/z 316.

A solution of 6-(2-methoxy-3-o-tolylureido)pyridylacetic acid (1.61 g,5.10 mmol) in DMF at 0° C. was treated with EDC (1.00 g, 5.2 mmol). Themixture was stirred at 0° C. for 1 to 2 h and then N-hydroxysuccinimide(0.60 g, 5.2 mmol) was added. After stirring at RT for more than 3 h,the reaction mixture was poured into 60% sat. NaHCO₃ and theN-succinimidyl 6-(2-methoxy-3-o-tolylureido)pyridylacetate filtered.

H-LD(OBn)V-NHCH₃:

H-LD(OBn)V-NHCH₃ was prepared by sequentially using procedure B withBOC-Val-OSu and methylamine, procedure D, procedure B withBOC-Asp(OBn)-OSu, procedure D, procedure B with BOC-Leu-OSu, thenprocedure D.

H-LD(OBn)V-OCH₃:

H-LD(OBn)V-OCH₃ was prepared by sequentially using procedure B withBOC-Asp(OBn)-OSu and H-Val-OMe, procedure D, procedure B withBOC-Leu-OSu, then procedure D.

H-LD(OBn)V-OBn:

H-LD(OBn)V-OBn was prepared by sequentially using procedure B withBOC-Asp(OBn)-OSu and H-Val-OBn, procedure D, procedure B withBOC-Leu-OSu, then procedure D.

H-LD(OBn)VP-OBn:

H-LD(OBn)VP-OBn was prepared by sequentially using procedure B withBOC-Val-OSu and H-Pro-OBn, procedure D, procedure B withBOC-Asp(OBn)-OSu, procedure D, procedure B with BOC-Leu-OSu, thenprocedure D.

H-LD(OBn)VP-OMe:

H-LD(OBn)VP-OMe was prepared by sequentially using procedure A withBOC-Val-OH and H-Pro-OMe, procedure,D, procedure B withBOC-Asp(OBn)-OSu, procedure D, procedure B with BOC-Leu-OSu, thenprocedure D.

H-LDVP-OH:

H-LDVP-OH was prepared by sequentially using procedure B withBOC-Val-OSu and H-Pro-OBn, procedure D, procedure B withBOC-Asp(OBn)-OSu, procedure D, procedure B with BOC-Leu-OSu, procedureF, then procedure D.

H-MD (OBn)VP-OBn:

H-MD(OBn)VP-OBn was prepared by sequentially using procedure B withBOC-Val-OSu and H-Pro-OBn, procedure D, procedure B withBOC-Asp(OBn)-OSu, procedure D, procedure B with BOC-Met-OSu, thenprocedure D.

H-LD(OBn)VP-NH₂:

H-LD(OBn)VP-NH₂ was prepared by sequentially using procedure B withBOC-Val-OSu and H-Pro-NH₂, procedure D, procedure B withBOC-Asp(OBn)-OSu, procedure D, procedure B with BOC-Leu-OSu, thenprocedure D.

Resin (MBC1):

Modified resin MBC1 (0.437 mmol/g) was synthesized according to theliterature procedure (see: Richter, L. S., et al., Tetrahedron Lett. 35,p. 5547 (1994)). MBC1 was treated with 50% TFA/CH₂Cl₂ and triethylsilanefor 2 h at RT then washed with CH₂Cl₂ (2×), isopropanol (1×), and CH₂Cl₂(3×) before use.

MBC2:

MBC2 was prepared by sequentially using procedure G withBOC-Asp(OBn)-OH, procedure I, procedure G with BOC-Leu-OH, procedure I,then procedure G with 4-phenylureidophenylacetic acid.

MBC3:

MBC3 was prepared by sequentially using procedure G with BOC-Val-OH,procedure I, procedure G with BOC-Asp(OBn)-OH, procedure I, procedure Gwith BOC-Leu-OH, procedure I, then procedure G with4-phenylureidophenylacetic acid.

MBC4:

MBC4 was prepared by sequentially using procedure G with BOC-Pro-OH,procedure I, procedure G with BOC-Val-OH, procedure I, procedure G withBOC-Asp(OBn)-OH, procedure I, procedure G with BOC-Leu-OH, procedure I,then procedure G with 4-phenylureidophenylacetic acid.

Example 2

Compound 77:

Compound 77 was prepared by using procedure A with picolinic acid andH-LD(OBn)V-OBn then procedure F. Purification by HPLC gave the titlecompound: m/z 451.

Example 3

Compound 64:

Compound 64 was prepared by using procedure A with hydrocinnamic acidand H-LD(OBn)V-OBn then procedure F. Purification by HPLC gave the titlecompound: m/z 478.

Example 4

Compound 155:

Compound 155 was prepared by using procedure B with chloro4-phenylbutyrate and H-LD(OBn)V-OBn then procedure F. Purification byHPLC gave the title compound: m/z 492.

Example 5

Compound 157:

Compound 157 was prepared by using procedure B with BOC-Asp(OBn)-OSu andisobutylamine, procedure D, procedure B with BOC-Leu-OSu, procedure D,procedure B with iQn-OSu, then procedure F. Purification by HPLC gavethe title compound: m/z 457.

Example 6

Compound 164:

Compound 164 was prepared by using procedure B with Qn-OSu andH-LD(OBn)V-NHCH₃ then procedure F. Purification by HPLC gave the titlecompound: m/z 514.

Example 7

Compound 174:

Compound 174 was prepared by using procedure B with BOC-Asp(OBn)-OSu andvalinol, procedure D, procedure B with BOC-Leu-OSu, procedure D,procedure B with Qn-OSu, then procedure F. Purification by HPLC gave thetitle compound: m/z 487.

Example 8

Compound 177:

Compound 177 was prepared by using procedure B with BOC-Asp(OBn)-OSu andH-Thr-OCH₃, procedure D, procedure B with BOC-Leu-OSu, procedure D,procedure B with 4-methoxybenzenesulfonyl chloride, then procedure F.Purification by HPLC gave the title compound: m/z 532.

Example 9

Compound 180:

Compound 180 was prepared by using procedure B with BOC-Val-OSu andmethylamine, procedure D, procedure B with BOC-Asp(OBn)-OSu, procedureD, procedure B with BOC-N-MeLeu-OSu, procedure D, procedure B withphenylacetyl chloride, then procedure F. Purification by HPLC gave thetitle compound: m/z 491.

Example 10

Compound 189:

Compound 189 was prepared by using procedure B with BOC-Val-OSu andmethylamine, procedure D, procedure B with BOC-Asp(OBn)-OSu, procedureD, procedure B with BOC-Leu-OSu, procedure D, procedure B withphenylsulfonyl chloride, then procedure F. Purification by HPLC gave thetitle compound: m/z 499.

Example 11

Compound 345:

Compound 345 was prepared by using procedure A with4-o-tolylureidophenylacetic acid and H-LD(OBn)V-OBn then procedure F.Purification by HPLC gave the title compound: m/z 606.

Example 12

Compound 206:

Compound 206 was prepared by using procedure A with4-o-tolylureidophenylacetic acid and H-LD(OBn)VP-OBn then procedure F.Purification by HPLC gave the title compound: m/z 709.

Example 13

Compound 144:

Compoound 144 was prepared by using procedure A with4-(2-hydroxyphenylureido) phenylacetic acid and H-LD(OBn)VP-OBn thenprocedure F. Purification by HPLC gave the title compound: m/z 711, 24.6min (gradient 8).

Example 14

Compound 145:

Compound 145 was prepared by using procedure A with4-phenylureidophenylacetic acid and H-LD(OBn)VP-OBn then procedure F.Purification by HPLC gave the title compound: m/z 695, 26.8 min(gradient 8).

Example 15

Compound 146:

Compound 146 was prepared by using procedure A with4-(2-hydroxyphenylureido) phenylacetic acid and H-MD(OBn)VP-OBn thenprocedure F. Purification by HPLC gave the title compound: m/z 729, 22.4m in (gradient 8).

Example 16

Compound 1:

Compound 1 was prepared by using procedure A with3-methoxy-4-phenylureidophenylacetic acid and H-LD(OBn)VP-OBn thenprocedure F. Purification by HPLC gave the title compound: m/z 725, 28.5min (gradient 8).

Example 17

Compound 2:

Compound 2 was prepared by using procedure A with3-methoxy-4-phenylureidophenylacetic acid and H-MD(OBn)VP-OBn thenprocedure F. Purification by HPLC gave the title compound: m/z 743, 27.0min (gradient 8).

Example 18

Compound 315:

Compound 315 was prepared by using procedure A with4-o-tolylureidophenylacetic acid and H-MD(OBn)VP-OBn then procedure F.Purification by HPLC gave the title compound: m/z 727.

Example 19

Compound 346:

Compound 346 was prepared by using procedure B withN-Hydroxysuccinimidyl 4-(2-(3-methylpyridylureido)phenylacetate andH-LDVP-OH. Purification by HPLC gave Compound 346: m/z 710.

Example 20

Compound 316:

Compound 316 was prepared by using procedure B withN-hydroxysuccinyimidyl 4-(2-pyridylureido)phenylacetate and H-LDVP-OH.Purification by HPLC gave the title compound: m/z 696.

Example 21

Compound 4:

Compound 4 was prepared by using procedure B with N-hydroxysuccinimidyl6-(2-methoxy-3-o-tolylureido)pyridylacetate and H-LDVP-OH. Purificationby HPLC gave the title compound: m/z 740, 30.7 min (gradient 8).

Example 22

Compound 147:

Compound 147 was prepared by using procedure B withN-hydroxysuccinimidyl 3-methoxy-4-phenylureidophenylacetate andH-LD(OBn)VP-NH₂ then procedure F. Purification by HPLC gave the titlecompound: m/z 724, 26.7 min (gradient 8).

Example 23

Compound 148:

Compound 148 was prepared by using procedure A with4-o-tolylureidophenylacetic acid and H-LD(OBn)VP-NH₂ then procedure F.Purification by HPLC gave the title compound: m/z 708, 26.0 min(gradient 8).

Example 24

Compound 317:

Compound 317 was prepared by using procedure B withN-hydroxysuccinimidyl 6-(2-methoxy-3-o-tolylureido)pyridylacetate andH-LD(OBn)VP-NH₂ then procedure F. Purification by HPLC gave the titlecompound: m/z 739, 28.0 min (gradient 8).

Example 25

Compound 336:

Compound 336 was prepared by using procedure A with4-(2-fluorophenyl)ureidophenylacetic acid and H-LD(OBn)VP-OBn thenprocedure F. Purification by HPLC gave the title compound: m/z 713.

Example 26

Compound 32:

Compound 32 was prepared by using procedure B with iQn-OSu andH-LD(OBn)VP-OBn then procedure F. Purification by HPLC gave the titlecompound: m/z 598, 24.7 min (gradient 8).

Example 27

Compound 34:

Compound 34 was prepared by using procedure B with phenylacetyl chlorideand H-LD(OBn)VP-OBn then procedure F. Purification by HPLC gave thetitle compound: m/z 561, 23.7 min (gradient 8).

Example 28

Compound 39:

Compound 39 was prepared by using procedure A with3-(-4-hydroxyphenyl)propionic acid and H-LD(OBn)VP-OMe then procedure F.Purification by HPLC gave Compound 39: m/z 591, 21.5 min (gradient 8).

Example 29

Compound 42:

Crude compound 42 was prepared by sequentially using procedure A withBOC-Val-OH and H-homoPro-OBn, procedure D, procedure B withBOC-Asp(OBn)-OSu, procedure D, procedure B with BOC-Leu-OSu, procedureD, procedure B with phenyl acetyl chloride then procedure F.Purification by HPLC gave the title compound: m/z 575, 26.4 min(gradient 8).

Example 30

Compound 52:

Compound 52 was prepared by sequentially using procedure A withBOC-norVal-OH and methylamine, procedure D, procedure B withBOC-Asp(OBn)-OSu, procedure D, procedure B with BOC-Leu-OSu, procedureD, procedure B with Qn-OSu then procedure F. Purification by HPLC gavethe title compound: m/z 518, 30.2 min (gradient 8).

Example 31

Compound 46:

Compound 46 was prepared by sequentially using procedure A withBOC-Val-OH and methylamine, procedure D, procedure B withBOC-Asp(OBn)-OSu, procedure D, procedure A with BOC-N-MeLeu-OH,procedure D, procedure A with 3-(4-hydroxyphenyl)propionic acid thenprocedure F. Purification by HPLC gave the title compound: m/z 521, 18.7min (gradient 8).

Example 32

Compound 61:

Compound 61 was prepared by sequentially using procedure B withBOC-Thr-OSu and morpholine, procedure D, procedure B withBOC-Asp(OBn)-OSu, procedure D, procedure B with BOC-Leu-OSu, procedureD, procedure B with Qn-OSu then procedure F. Purification by HPLC gavethe title compound: m/z 572, 24.0 min (gradient 8).

Example 33

Compound 213:

Compound 213 was prepared by using procedure H with MBC2 and benzyaminethen procedure J: m/z 588.

Example 34

Compound 214:

Compound 214 was prepared by using procedure H with MBC2 and morpholinethen procedure J: m/z 568.

Example 35

Compound 215:

Compound 215 was prepared by using procedure H with MBC2 andisopropylamine then procedure J: m/z 540.

Example 36

Compound 216:

Compound 216 was prepared by using procedure H with MBC2 andcyclohexylamine then procedure J: m/z 580.

Example 37

Compound 217:

Compound 217 was prepared by using procedure H with MBC2 andisobutylamine then procedure J: m/z 554.

Example 38

Compound 218:

Compound 218 was prepared by using procedure H with MBC2 and piperdinethen procedure J: m/z 566.

Example 39

Compound 318:

Compound 318 was prepared by using procedure H with MBC3 and morpholinethen procedure J: m/z 667.

Example 40

Compound 319:

Compound 319 was prepared by using procedure H with MBC3 andisopropylamine then procedure J: m/z 640.

Example 41

Compound 320:

Compound 320 was prepared by using procedure H with MBC3 andcyclohexylamine then procedure J: m/z 679.

Example 42

Compound 321:

Compound 321 was prepared by using procedure H with MBC3 and benzylaminethen procedure J: m/z 687.

Example 43

Compound 322:

Compound 322 was prepared by using procedure H with MBC3 and piperidinethen procedure J: m/z 665.

Example 44

Compound 323:

Compound 323 was prepared by using procedure H with MBC3 andisobutylamine then procedure J: m/z 653.

Example 45

Compound 324:

Compound 324 was prepared by using procedure H with MBC4 andcyclohexylamine then procedure J: m/z 777.

Example 46

Compound 325:

Compound 325 was prepared by using procedure H with MBC4 and piperdinethen procedure J: m/z 763.

Example 47

Compound 326:

Compound 326 was prepared by using procedure H with MBC4 and benzylaminethen procedure J: m/z 785.

Example 48

Compound 327:

Compound 327 was prepared by using procedure H with MBC4 andisopropylamine then procedure J: m/z 736.

Example 49

Compound 328:

Compound 328 was prepared by using procedure H with MBC4 andisobutylamine then procedure J: m/z 750.

Example 50

Compound 363

A. A mixture of o-tolylureidophenylacetic acid (3.53 g, 12.4 mmol),H-Leu-OtBu.HCl (2.78 g, 12.4 mmol), TBTU (3.98 g, 12.4 mmol), andiPr₂NEt (4.32 mL, 24.8 mmmol) in DMF (25 mL) was stirred overnight atRT. The product was precipitated by addition of H2O (10 mL). The solidswere collected by filtration on a medium frit, washing with 2:1 DMF/H₂O(35 mL), H₂O (25 mL), and Et₂O (2×25 mL), and dried on the filter (4.18g, 74%). All of this product was suspended in CH₂Cl₂ (16 mL) and treatedwith TFA (16 mL) and stirred at RT 2 hr. The reaction was concentratedto a syrup which was evaporated from CH₂Cl₂ (2×20 mL). The residue wastriturated with Et₂O (100 mL) at RT for 2 hr. The solids were collectedby filtration on a medium frit, washing with Et₂O (50 mL), and dried onthe filter (3.40 g, 93%): MS (FAB) 398.

B. A mixture of DCC (0.206 g, 1.0 mmol) and HOBT (0.135 g, 1.0 mmol) inEtOAc (6 mL) was stirred at RT 20 min until homogeneous. Fmoc-Asp-OtBu(0.411 g, 1.0 mmol), piperonylamine (0.12 mL, 1.0 mmol), andN-methylmorpholine (0.22 mL, 2.0 mmol) were added. After stirringovernight, the reaction was filtered to remove solids and the cakewashed with fresh EtOAc (10 mL). The filtrate was washed with H₂O (2×),5% citric acid (1×), 5% NaHCO₃ (1×), and brine (1×), and dried (MgSO₄).Flash column chromatography on SiO₂ eluting with 100% CHCl₃ to 2%MeOH/CHCl₃ provided 0.54 g (100%) of pure product as a solid:mp=128-130° C.; TLC(2% MeOH/CHCl₃) R_(f)=0.10; MS (FAB) 545; ¹H NMR(CDCl₃, 300 MHz, ppm) 7.75-7.72 (m, 2H), 7.59-7.56 (m, 2H), 7.40-7.34(m, 2H), 7.30-7.25 (m, 2H), 6.71-6.66 (m, 3H), 6.13-6.10 (m, 2H), 5.84(s, 2H), 4.46 (m, 1H), 4.38-4.16 (m, 5H), 2.86 (dd, 1H, J=4.7, 15.6 Hz),2.72 (dd, 1H, J=4.16, 15.6 Hz), 1.45 (s, 9H).

C. The product from Example 50B (0.25 g, 0.46 mmol), piperidine (0.45mL, 4.6 mmol), and CH₂Cl₂ (0.45 mL) were stirred at RT for 90 min. Thereaction was evaporated to a solid residue. Flash column chromatographyon SiO₂ using a MeOH/EtOAc gradient provided product (0.138 g, 93%) as acolorless oil: MS (FAB) 323; TLC(10% MeOH/EtOAc) R_(f)=0.15; ¹H NMR(CDCl₃, 300 MHz, ppm) 7.63 (br s, 1H), 6.75-6.68 (m, 3H), 5.90 (s, 2H),4.34 (dd, 1H, J=5.7, 14.7 Hz), 4.28 (dd, 1H, J=5.7, 14.7 Hz), 3.65 (dd,1H, J=3.4, 9.3 Hz), 2.62 (dd, 1H, J=3.4, 15.7 Hz), 2.38 (dd, 1H, J=9.3,15.7 Hz), 1.74 (s, 2H), 1.42 (s, 9H).

D. The product from Example 50C (2.55 g, 7.91 mmol) and Eschenmoser'ssalt (1.61 g, 8.70 mmol) were refluxed in MeCN (80 mL) under an inertatmosphere for 42 hr. The reaction was cooled to RT and evaporated todryness. The residue was diluted with 5% NaHCO₃ and extracted with EtOAc(3×). The combined organic extracts were washed with 5% NaHCO₃ (1×), H₂O(1×), and brine (1×), and dried (MgSO₄). The crude product was dissolvedin Et₂O (250 mL) and passed through a short pad of SiO₂, eluting withEt₂O followed by EtOAc. The slightly impure product thus obtained wasfurther purified by trituration with ice cold Et₂O (30 mL) and collectedby filtration to give a white solid (0.904 g, 34%): mp=121-123° C.; TLC(10% MeOH/CHCl₃) R_(f)=0.59; ¹H NMR (CDCl₃, 300 MHz, ppm) 6.75-6.66 (m,3H), 5.92 (s, 2H), 4.66 (A of AB, 1H, J=14.7 Hz), 4.23 (B of AB, 1H,J=14.7 Hz), 4.15 (ABq, 2H, J=11.9 Hz), 3.68 (dd, dd, 1H, J=5.2, 10.9Hz), 2.72 (dd, 1H, J=5.2, 17.3 Hz), 2.41 (dd, 1H, J=10.9, 17.3 Hz), 1.45(s, 9H); C,H,N for C₁₇H₂₂N₂O₅, theory—C: 61.07, H: 6.63, N: 8.38,found—C: 60.80, H: 6.59, N: 8.22.

E. The product from Example 50D (0.50 g, 1.5 mmol), the product fromExample 50A (0.596 g, 1.5 mmol), and EDC (0.314 q, 1.64 mmol) werestirred in NMP (3 mL) at RT for 48 hr. The reaction was poured intoEtOAc (60 mL), washed with H₂O (8×6 mL), brine (1×), and dried (MgSO₄).Flash column chromatography on SiO₂ eluting with 100% CHCl₃ to 30%EtOAc/CHCl₃ provided product (0.94 g, 88%) as a pale yellow oil: MS(FAB) 714; TLC (10% MeOH/CHCl₃) R_(f)=0.40; ¹H NMR (CDCl₃, 300 MHz, ppm)consistent with structure and indicative of diastereomers.

F. The product from Example 50E (0.94 g, 1.32 mmol) was stirred in TFA(10 mL) at RT for 3 hr. The reaction was concentrated to dryness and theresidue evaporated from CH₂Cl₂ (3×10 mL). The crude product wastriturated with Et₂O at RT, collected by filtration and dried on thefilter (0.733 g, 84%): MS (FAB) 658 (M+H), 680 (M+Na); TLC (5%HOAc/EtOAc) R_(f)=0.15; ¹H NMR (d⁶-DMSO, 300 MHz, ppm) consistent withstructure and indicative of diastereomers.

Example 51

Compound 364:

A. In the same manner as described in Example 50B, Fmoc-Asp-OtBu (8.23g, 20.0 mmol) was reacted with H-Gly-OBn.HCl (4.03 g, 20.0 mmol). Flashcolumn chromatography on SiO₂, using an EtOAc/hexane gradient providedproduct (9.8 g, 88%) as a waxy solid: MS (FAB) 559; TLC (10% MeOH/CHCl₃)R_(f)=0.71;

¹H NMR (CDCl₃, 300 MHz, ppm) 7.73 (d, 2H, J=7.5 Hz), 7.59 (d, 2H, J=7.4Hz), 7.40-7.26 (m, 9H), 6.44 (br s, 1H), 6.09 (d, 1H, J=8.3 Hz), 5.13(s, 2H), 4.52-4.49 (m, 1H), 4.41-4.29 (m, 2H), 4.21 (t, 1H, J=7.1 Hz),4.04 (d, 2H, J=5.2 Hz), 2.95 (dd, 1H, J=4.6, 15.7 Hz), 2.79 (dd, 1H,J=4.3, 15.7 Hz), 1.46 (s, 9H).

B. The product of Example 51A (9.8 g, 17.54 mmol) was deprotected in themanner described in Example 50C. Filtration through a pad of SiO₂ with100% EtOAc followed by 5% MeOH/CHCl₃ provided product (4.24 g, 72%) asan oil: MS (FAB) 337; TLC (3% MeOH/EtOAc) R_(f)=0.15; ¹H NMR (CDCl₃, 300MHz, ppm) 8.00 (t, 1H, J=5.4 Hz), 7.30-7.21 (m, 5H), 5.07 (s, 2H), 3.98(AB of ABX, 2H, J=5.4, 18.1 Hz), 3.60 (dd, 1H, J=3.4, 9.2 Hz), 2.60 (dd,1H, J=3.4, 5.4 Hz), 2.38 (dd, 1H, J=9.2, 15.4 Hz), 1.79 (br s, 2H), 1.36(s, 9H).

C. The product of Example 51B (4.24 g, 12.60 mmol) was cyclized in themanner described in Example 50D. Flash column chromatography using anEtOAc/CHCl₃gradient provided product as a syrup (1.4 g, 32%): MS (FAB)349; TLC (1:1 EtOAc/CHCl₃) R_(f)=0.53; ¹H NMR (CDCl₃, 300 MHz, ppm)7.35-7.25 (m, 5H), 5.11 (s, 2H), 4.21 (A of AB, 1H, J=17.5 Hz), 3.95 (Bof AB, 1H, J=17.5 Hz), 3.71 (dd, 1H, J=5.1, 11.2 Hz), 2.68 (dd, 1H,J=5.1, 17.2 Hz), 2.36 (dd, 1H, J=11.2, 17.2 Hz), 1.43 (s, 9H).

D. The product of Example 51C (1.40 g, 4.02 mmol) was coupled with theproduct of Example 50A using the procedure of Example 50E. Flash columnchromatography using a CHCl₃/EtOAc gradient provided product as abrittle, pale yellow foam (2.21 g, 76%): MS (FAB) 728; TLC (1:1CHCl₃/EtOAc) R_(f)=0.28; ¹H NMR (CDCl₃, 300 MHz, ppm) consistent withstructure and indicative of diastereomers.

E. The product of Example 51D (0.15 g, 0.21 mmol) was deprotected andpurified as described in Example 50F. The product was obtained as anoff-white solid (0.127 g, 90%): MS (FAB) 672 (M+H), 695 (M+Na); TLC(9:1:0.1 CHCl₃/MeOH/AcOH) Rf=0.54; ¹H NMR (d⁶-DMSO, 300 MHz, ppm)consistent with structure and indicative of diastereomers.

Example 52

Compound 365

A. The product from Example 51E (0.100 g, 0.15 mmol),4-methoxybenzylamine (20 μL, 0.15 mmol), and TBTU (0.0482 g, 0.15 mmol)in NMP (0.3 mL) were treated with iPr₂NEt (78 μL, 0.45 mmol). Afterstirring overnight at RT, the reaction was diluted with EtOAc (10 mL),washed with H₂O (5×2 mL), 5% citric acid (2×2 mL), 5% NaHCO₃ (2×2 mL)and brine (1×2 mL) and dried (MgSO₄). Filtration through a short pad ofSiO₂, eluting with 2% MeOH/CHCl₃ followed by 4% MeOH/CHCl₃, providedproduct as a foam (0.087 g, 73%): MS (FAB) 792; TLC (9:1 CHCl₃/MeOH)R_(f)=0.41; ¹H NMR (CDCl₃, 300 MHz, ppm) consistent with structure andindicative of diastereomers.

B. A suspension of the product of Example 52A (0.087 g, 0.11 mmol) andDegussa type E101 NE/W 10% Pd/C (0.017 g) in MeOH (10 mL) washydrogenated under 25 psi H₂ for 18 hr. The reaction was filteredthrough Celite, rinsing with MeOH. The filtrate was evaporated todryness. The residue was triturated with Et₂O and the resultant beigesolids collected by filtration (36.1 mg, 47%): MS (FAB) 701 (M+H), 723(M+Na); ¹H NMR (d⁶-DMSO, 300 MHz, ppm) consistent with structure andindicative of diastereomers.

Example 53 Inhibition of VLA4-dependent Adhesion to BSA-CS1

This assay was used to assess the potency of VLA4-directed inhibitorycompounds of this invention.

1. Conjugation of CS1 to BSA

We dissolved BSA-SMCC (Pierce Chemical, Rockford, Ill.; Catalog #77115)in H₂O at a concentration of 10 mg/mL. [SEQ ID NO:4]:Cys-Tyr-Asp-Glu-Leu-Pro-Gln-Leu-Val-Thr-Leu-Pro-His-Pro-Asn-Leu-His-Gly-Pro-Glu-Ile-Leu-Asp-Val-Pro-Ser-Thr(“Cys-Tyr-CS1 peptide”), which we synthesized by conventional solidphase chemistry and purified by HPLC, was dissolved in 10 mM HEPES pH 5,50 mM NaCl and 0.1 mM EDTA also at a concentration of 10 mg/mL. We thenmixed 500 μL of BSA-SMCC, 250 μL of Cys-Tyr-CS1 peptide and 75 μL of 1mM HEPES pH 7.5 and allowed the conjugation reaction to proceed for 30minutes. We stopped the reaction by adding 1 μL of beta-mercaptoethanol.Samples were analyzed for cross-linking by SDS-PAGE. This reactionproduced multiple molecules of the Cys-Tyr-CS1 peptide conjugate to eachBSA molecule.

2. Preparation of Plates for Adhesion Assay

We coated the wells of a Linbro titertek polystyrene 96-well flat bottomplate (Flow Laboratories, Maclean, Va.; catalog #76-231-05) with 100 μLof the above-described BSA-CS1 solution diluted to 1 μg/mL in 0.05 MNaHCO₃ (15 mM NaHCO₃, 35 mM Na₂CO₃) pH 9.2. Some wells were not coatedwith CS1 in order to assess non-specific cell binding (NSB). The platewas then incubated overnight at 4° C.

Following this incubation, the contents of the wells were removed byinverting and blotting the plate. All of the wells were then blockedwith 100 μL of 1% BSA in PBS, 0.02% NaN₃, for a minimum of one hour atroom temperature.

3. Preparation of Fluorescently Labelled Ramos Cells

Ramos cells are grown, maintained and labelled in RPMI 1640 culturemedium containing 1% BSA. Just prior to running the assay, we added2′,7′-bis-(2-carboxyethyl)-5 (and -6) carboxyfluorescein acetoxymethylester (“BCECF-AM”; Molecular Probes Inc., Eugene, Oreg.; catalog#B-1150) to a final concentration of 2 μM to a culture of Ramos cells(4×10⁶ cells/mL). We incubated the cells for 20 minutes at 37° C.

Following labelling, the cells were washed twice in assay buffer (24 mMTRIS, 137 mM NaCl, 2.7 mM KCl, pH 7.4, containing 0.1% BSA and 2 mMglucose) to remove any cations originating from the culture medium. Thecells were then resuspended in assay buffer to 4×10⁶ cells/mL and 2 mMMnCl₂ was added to upregulate VLA4 on the surface of the cells.

4. Running the Assay

Immediately prior to running the assay, we removed the BSA blockingsolution from the 96-well plates and washed the wells with 100 μL ofassay buffer. We then added to each well 25 μL of test cell adhesioninhibitory compound at 2× the final concentration and 25 μL of thelabelled Ramos cells. Final concentrations were selected across a rangeof anticipated IC50s, usually between 0.01 nM-10 μM. Each concentrationof compound was tested in triplicate. The compound and cells are allowedto incubate for 30 minutes at room temperature.

We then emptied the contents of the plate and washed the wells 4 timeswith assay buffer. Using a light microscope, we examined the NSB wells.If more than a few cells are bound to those wells, we washed the plateonce more to remove the excess non-specifically bound cells.

Binding of the Ramos cells to the CS1 peptide-coated wells was measuredby adding 100 μL of assay buffer to each well and quantitatingfluorescence in a Millipore Cytofluor 2300 System plate reader set at485 nm excitation and 530 nm emission. Binding was expressed as anIC50—the concentration of inhibitor at which 50% of control bindingoccurs. Percent binding is calculated by the formula:[(F _(TB) −F _(NS))−(F _(I) −F _(NS))]/[(F _(TB) −F _(NS))×100=%binding,where F_(TB) is total fluorescence bound to CS1-containing wells withoutadded inhibitor; F_(NS) is fluorescence bound in wells lacking CS1; andF_(I) is fluorescence bound in wells containing an inhibitor of thisinvention.

Other compounds according to this invention were similarly assayed. TheIC50 range for each of these compounds is indicated in the table below:

Cmpd # IC₅₀ 1 A 2 A 3 A 4 A 5 C 6 C 7 C 8 C 9 C 10 C 11 C 12 C 13 B 14 C15 C 16 C 17 C 18 B 19 C 20 C 21 C 22 C 23 C 24 C 25 C 26 C 27 C 28 C 29C 30 C 31 C 32 C 33 C 34 B 35 B 36 C 37 C 38 C 39 C 40 C 41 B 42 B 43 B44 B 45 C 46 C 47 C 48 C 49 C 50 C 51 C 52 C 53 C 54 C 55 C 56 B 57 B 58B 59 B 60 C 61 B 62 C 63 C 64 C 65 C 66 C 67 C 68 C 69 C 70 C 71 C 72 C73 C 74 C 75 C 76 C 77 C 78 C 79 C 80 C 81 C 82 C 83 C 84 C 85 C 86 C 87C 88 C 89 C 90 C 91 C 92 C 93 C 94 C 95 C 96 C 97 C 98 C 99 C 100 C 101C 102 C 103 C 104 C 105 C 106 C 107 C 108 C 109 C 110 C 111 C 112 C 113C 114 C 115 C 116 C 117 C 118 C 119 C 120 C 121 C 122 C 123 C 124 C 125C 126 C 127 C 128 C 129 C 130 C 131 C 132 C 133 C 134 C 135 C 136 C 137C 138 C 139 C 140 C 141 C 142 C 143 C 144 A 145 A 146 A 147 A 148 A 149C 150 C 151 C 152 C 153 C 154 C 155 C 156 B 157 B 158 C 159 C 160 C 161C 162 C 163 C 164 B 165 C 166 C 167 C 168 B 169 C 170 C 171 C 172 C 173C 174 B 175 B 176 C 177 C 178 C 179 C 180 B 181 C 182 C 183 C 184 C 185C 186 C 187 C 188 C 189 C 190 C 191 C 192 B 193 C 194 nd 195 C 196 C 197B 198 C 199 B 200 B 201 B 202 B 203 A 204 B 205 B 206 A 207 B 208 A 209B 210 B 211 B 212 B 213 B 214 B 215 B 216 B 217 B 218 B 219 A 220 A 221A 222 A 223 A 224 A 225 A 226 A 227 A 228 A 229 A 230 A 231 A 232 A 233A 234 A 235 A 236 A 237 A 238 A 239 A 240 A 241 A 242 A 243 A 244 A 245A 246 A 247 A 248 A 249 A 250 A 251 A 252 A 253 A 254 A 255 A 256 A 257A 258 A 259 A 260 A 261 A 262 A 263 A 264 A 265 A 266 A 267 A 268 A 269A 270 A 271 A 272 A 273 A 274 A 275 A 276 A 277 A 278 A 279 A 280 A 281A 282 A 283 A 284 A 285 A 286 A 287 A 288 A 289 A 290 A 291 A 292 A 293A 294 A 295 A 296 A 297 A 298 A 299 A 300 A 301 A 302 A 303 A 305 A 306A 305 A 307 A 308 A 309 A 310 A 311 A 312 A 313 A 314 A 315 A 316 A 317A 318 A 319 B 320 B 321 B 322 A 323 B 324 B 325 B 326 A 327 A 328 A 329A 330 C 331 C 332 C 333 C 334 C 335 C 336 A 337 A 338 A 339 C 340 C 341C 342 C 343 C 344 C 345 A 346 A 347 A 348 C 349 C 350 C 351 C 352 C 353C 354 C 355 nd 356 nd 357 nd 358 A 359 A 360 C 361 C 362 C 363 C 364 B365 B 366 C 367 C 368 C 369 C 370 C 371 B 372 C 373 C 374 C 375 B 376 C377 nd 378 nd 379 nd 380 C 381 nd 382 nd 383 nd 384 nd 385 nd 386 C 387C 388 nd 389 nd 390 C 391 C 392 nd 393 C 394 C 395 C 396 nd 398 C 399 nd400 nd 401 nd 402 nd 403 C 404 C 405 C 406 C 407 C 408 C 409 B 410 B 411A 412 B 413 B 414 B 415 B 416 B Table abbreviations: A — <50 nm; B — 50nm-10μm; C — >10 μm; nd — not determined. All compounds tested in thistable demonstrated an IC₅₀ < 1 mM.

Example 54 Direct Binding of VLA4-Presenting Cells to VCAM-IgG

We next examined the ability of the compounds of this invention toinhibit VCAM/VLA4 binding, utilizing a VCAM-IgG-alkaline phosphataseconjugate. To carry out this assay, we used the Millipore MultiscreenAssay System (Millipore Corp., Bedford, Mass.) to wash the cellsefficiently.

1. Preparation of VCAM-IgG-AP Conjugates

The construction of VCAM 2D-IgG expression vectors, transfection of CHOcells with those constructs and purification of the resulting expressionproduct is described in PCT publication WO 90/13300, the disclosure ofwhich is herein incorporated by reference.

1.2 ml of purified VCAM 2D-IgG (5 mg/ml in 10 mM HEPES, pH 7.5) wasreacted with 44 μl of Traut's reagent (2-iminothiolane, 20 mg/ml inwater; Pierce Chemical, Rockford, Ill.) at room temperature for 30minutes. The sameple was desalted on a 15 ml Sephadex G-25 columnequilibrated with 100 mM NaCl, 10 mM MES, pH 5.0. One ml fractions werecollected and absorbance at 280 nm was determined. The two peakfractions were pooled.

One ml of calf intestinal alkaline phosphatase (19 mg/ml; PierceChemical, Rockford, Ill.) was reacted with 100 μl of sulfo-SMCC (30mg/ml in water) and 100 μl 1 M HEPES, pH 7.5 for 35 minutes at roomtemperature. The reaction mix was desalted on a 12 ml Sephadex G-25column equilibrated with 150 mM NaCl, 10 mM HEPES, pH 6.0. One mlfractions were collected and absorbance at 280 nm was determined. Thetwo peak fractions were pooled and stored on ice.

The alkaline phosphatase-SMCC and VCAM 2D-IgG-iminothilane adducts werecross-linked at a molar ratio of 2:1 in Tris-HCL, pH 7.5 by incubationat room temperature for 30 minutes. Extent of cross-linking wasdetermined by SDS-PAGE. The cross-linked products were stabilized by theaddition of 2 mM MgCl₂ and 0.25 mM ZnCl₂ and stored at 4° C.

2. Binding Assay

We first blocked a 96-well filtration plate for by adding 275 μL of PBScontaining 0.1% Tween 20 and 2% BSA (“blocking buffer”) to each well andincubating for 1 hour at room temperature. The plate was then placedonto a vacuum manifold and the blocking buffer was drained through thebottom of the filtration wells into a waste collection tray. Then wewashed the wells three times with 200-250 μL of Tris-buffered saline,containing 0.1% BSA, 2 mM glucose and 1 mM HEPES, pH 7.5 (“assaybuffer”) to wash out any remaining blocking buffer. We then drained theplates and blotted them on paper towels to remove buffer on theunderside of the plate.

We then prepared a stock solution of VCAM-IgG-AP (4 μg/mL in assaybuffer) and filtered it through a 0.2 μ low protein binding syringefilter (Gelman Sciences, Ann Arbor, Mich. #4454). This solution was thendiluted 1:10 in assay buffer and 25 μL was added to every well of thewashed plate.

We diluted the cell adhesion inhibitor being tested to 2× finalconcentration in assay buffer and added 25 μL of each dilution totriplicate wells in the plate. Final concentrations used ranged from0.01 nM-10 μM. Control wells for total binding and non-specific bindingreceived 25 μL of assay buffer, instead of inhibitor. Total bindingwells contained cells and VCAM-IgG-AP in assay buffer. Non-specificbinding wells contained only VCAM-IgG-AP in assay buffer.

Jurkat cells were washed once in assay buffer to remove growth mediumand resuspended at 8×10⁶/mL in assay buffer containing 2 mM MnCl₂. Weadded 50 μl of Jurkat cells to every well, except the non-specificbinding wells, which received 50 μL of assay buffer to maintain a finalassay volume of 100 μL per well. We gently mixed the contents of thewells by tapping the sides of the plate. The plate was then allowed toincubate undisturbed for 60 minutes at room temperature.

At the end of the 60 minute incubation, we placed the plate on thevacuum manifold to drain the wells. We carefully added 100 μL of assaybuffer containing 1 mM MnCl₂ (wash buffer) to each well so as not todisturb the cells on the bottom. The wash buffer was removed by vacuumand the plate was washed again with 150 μL of wash buffer. Afterdraining the wash buffer again, the underside of the plate was blottedon paper towels.

Next, we prepared a 10 mg/mL solution of 4-nitrophenylphosphate in 0.1 Mglycine, 1 mM ZnCl₂, pH 10.5 (substrate buffer) and added 100 μLimmediately added to each well. The plate was incubated for 30 minutesat room temperature to allow the calorimetric reaction to proceed. Westopped the reaction by adding 100 μL of 3 N NaOH to each well.

The contents of the 96-well filtration plate was then transferreddirectly into a 96-well flat bottom plate using the vacuum manifold. Theplate was read at a wavelength of 405 nm to determine the amount of VCAMconjugate bound to the cells. Percent binding is calculated by theformula:[(A _(TB) −A _(NS))−(A ^(I) −A _(NS))]/[(A _(TB) −A _(NS))×100=%binding,where A_(TB) is the absorbance at 405 nm of CS1-containing wells withoutadded inhibitor; A_(NS) is the absorbance at 405 nm in wells lackingCS1; and A_(I) is absorbance at 405 nm in wells containing an inhibitorof this invention

We assayed other compounds of this invention in the same assay. The IC50values are comparable to those derived from the CS1 binding assaydescribed in the previous example, although certain compoundsdemonstrated up to 10-fold greater binding in this assay than in theprevious assay.

Example 55 Inhibition of Mouse Contact Hypersensitivity

We anesthetized 20-g female Balb/c mice (Jackson Laboratories, BarHarbor, Me.) with sodium pentobarbital (90 mg/kg, i.p.). A 3 cm² patchof abdominal skin was then exposed by closely shaving the fur. The skinwas then scrubbed with 70% ethanol, followed by application of 25 μL of0.5% DNFB in 4:1 v/v acetone:olive oil onto the bare abdominal skin. Wethen lightly scratched the skin with the applying pipet tip to encouragemild inflammation. Twenty four hours after the initial sensitization weagain sensitized the mouse with 25 μL of 0.5% DNFB at same abdominalskin location, again followed by light scratching with the pipet tip.The second sensitization was performed while restraining theunanesthetized mouse.

On Day 5 (120 hours after the initial sensitization), we anesthetizedthe mice with 90:10 mg/kg ketamine:xylazine, i.p. and applied asub-irritant dose of 10 μL of 0.2% DNFB to the dorsal surface of theleft ear. The right ear received a similar application of the 4:1 v/vacetone:olive oil vehicle.

Four hours after challenging the immune response, we administeredvarious concentrations of the inhibitors of this invention to the micein 100 μL 0.5% sodium phosphate buffer, pH 8.8, and 3% v/v DMSO bysubcutaneous (s.c.) injection. Less soluble inhibitors occasionallyrequired up to 30% DMSO addition the highest concentrations tested.Groups of 8 mice were used for each treatment tested. Positive (PS2anti-mouse VLA-4 antibody, 8 mg/kg, i.v.), and negative control(phosphate-buffered physiological saline, PBS, 100 μL i.v.; DMSO in PBS,100 μL s.c.) groups were routinely tested for comparison as part of theassay of test compounds.

Twenty four hours after challenge mice were again anesthetized withketamine:xylazine and the ear thickness of both ears measured with anengineer's micrometer to an accuracy of 10⁻⁴ inches. The ear swellingresponse for each mouse was the difference between its control- andDNFB-challenged ear thickness. Typical uninhibited ear swellingresponses were 65-75×10−4 in. Inhibition of the ear swelling responsewas judged by comparison of treated groups with their negative controlgroup. Percent inhibition was calculated as:$\left\lbrack {\frac{\begin{matrix}\left( {{mean}\quad{negative}\quad{control}\quad{group}} \right. \\\left. {{ear}\quad{swelling}} \right)\end{matrix} - \begin{matrix}\left( {{mean}\quad{test}\quad{group}\quad{ear}} \right. \\\left. {swelling} \right)\end{matrix}}{{mean}\quad{negative}\quad{control}\quad{group}\quad{ear}\quad{swelling}} \times 100} \right\rbrack$Statistical significance of the difference among treatment groups wasevaluated using one-way analysis of variance followed by computation ofthe Tukey-Kramer Honestly Significant Difference (JMP, SAS Institute)using p<0.05.

The inhibitors of this invention cause a statistically significantreduction in the ear swelling response of DNFB-treated mice as comparedto uninhibited control animals.

Example 56 Inhibition of Ascaris Antigen-induced Late Phase AirwaySensitivity in Allergic Sheep

Sheep which had previously been shown to develop both early and latebronchial responses to Ascaris suum antigen were used in this study. Theprotocol used for the experiment was that described in W. M. Abraham etal., J. Clin. Invest., 93, pp. 776-87 (1994), except that the VLA-4inhibitors of this invention were administered to the animals wasdissolved in 3-4 ml of 50% aqueous ethanol and delivered by aerosolspray.

The results showed that all of the VLA-4 inhibitors of this inventioninhibited the airway responses associated with administration of Ascarissuum antigen.

While we have hereinbefore presented a number of embodiments of thisinvention, it is apparent that our basic construction can be altered toprovide other compounds and methods which utilize the compounds of thisinvention. Therefore, it will be appreciated that the scope of thisinvention is to be defined by the claims appended hereto rather than thespecific embodiments which have been presented hereinbefore by way ofexample.

1. A cell adhesion inhibitory compound of formula (I),Z—(Y¹)—(Y²)—(Y³)_(n)—X  (I) or a pharmaceutically acceptable derivativethereof, wherein: Z is selected from the group consisting of aliphaticacyl substituted with N-arylamido; heterocycloyl; substitutedaralkylcarbonyl; heterocycloalkylcarbonyl; cycloalkylcarbonyl optionallyfused with aryl; heterocycloalkoxycarbonyl; alkylaminocarbonyl;arylamino-carbonyl optionally substituted with bis(alkylsulfonyl)amino,alkoxycarbonylamino or alkenyl; aralkylamino-carbonyl optionallysubstituted with bis(alkylsulfonyl)amino, alkoxycarbonylamino oralkenyl; alkylsulfonyl; aralkylsulfonyl; arylsulfonyl;cycloalkylsulfonyl optionally fused with aryl; heterocyclylsulfonyl;heterocyclylalkylsulfonyl; aryloxycarbonyl; cycloalkyloxycarbonyl;heterocyclyloxycarbonyl; heterocyclylalkoxycarbonyl; mono ordi-alkylaminocarbonyl optionally substituted with aryl;(alkyl)(aralkyl)aminocarbonyl; mono- or di-aralkylaminocarbonyl; mono-or di-arylaminocarbonyl; (aryl)(alkyl)aminocarbonyl; mono- ordi-cycloalkylaminocarbonyl; heterocyclylaminocarbonyl;heterocyclylalkylaminocarbonyl; (alkyl)(heterocyclyl)aminocarbonyl;(alkyl)(heterocyclylalkyl)aminocarbonyl;(aralkyl)(heterocyclyl)aminocarbonyl;(aralkyl)(heterocyclylalkyl)aminocarbonyl; alkenoyl optionallysubstituted with aryl; alkenylsulfonyl optionally substituted with aryl;alkynoyl optionally substituted with aryl; alkynylsulfonyl optionallysubstituted with aryl; cycloalkenylcarbonyl; cycloalkenylsulfonyl;cycloalkylalkylsulfonyl; arylaroyl; biarylsulfonyl; alkoxysulfonyl;aralkoxysulfonyl; alkylaminosulfonyl; aryloxysulfonyl;arylaminosulfonyl; N-arylurea-substituted alkanoyl;N-arylurea-substituted alkylsulfonyl; cycloalkenyl-substituted carbonyl;cycloalkenyl-substituted sulfonyl; alkenoxycarbonyl optionallysubstituted with aryl; alkenoxysulfonyl optionally substituted witharyl; alkynoxycarbonyl optionally substituted with aryl;alkynoxysulfonyl optionally substituted with aryl; alkynylaminocarbonyloptionally substituted with aryl; alkenylaminocarbonyl optionallysubstituted with aryl; alkenylaminosulfonyl optionally substituted witharyl; alkynylaminosulfonyl optionally substituted with aryl;acylamino-substituted alkanoyl; acylamino-substituted alkylsulfonyl;aminocarbonyl-substituted alkanoyl; carbamoyl-substituted alkanoyl;carbamoyl-substituted alkylsulfonyl; heterocyclylaminosulfonyl;carboxyalkyl-substituted aralkoyl; carboxyalkyl-substitutedaralkylsulfonyl; oxocarbocyclyl-fused aroyl; oxocarbocyclyl fusedarylsulfonyl; heterocyclylalkanoyl; N′,N′-alkyl arylhydrazinocarbonyl;aryloxy-substituted alkanoyl; and heterocyclylalkylsulfonyl; Y¹ is—N(R¹)—C(R²)(A¹)—C(O)—; Y² is —N(R¹)—C(R²)(A²)—C(O)—; each Y³ isrepresented by the formula —N(R¹)—C(R²)(A³)—C(O)—; A¹ is selected fromthe group consisting of amino acid side chains and correspondingprotected derivatives, cycloalkyl, and alkyl optionally substituted withamino, acylamino, amino-substituted acylamino, alkoxycarbonylamino,aryl, cycloalkyl, carboxy, alkoxy, aralkyloxy, alkoxycarbonyl,aralkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, (alkyl)(aralkyl)aminocarbonyl,aralkylaminocarbonyl, diaralkylaminocarbonyl, hydroxyl,carboxyalkylaminocarbonyl, hydroxylaminocarbonyl, mercapto, thioalkoxyor heterocycyl; A² is selected from the group consisting of acidicfunctional groups and alkyl optionally substituted with an acidicfunctional group, a protected acidic functional group or aryl; each A³,independently, is selected from the group consisting of amino acid sidechains and corresponding protected derivatives, aryl, cycloalkyl, andalkyl optionally substituted with amino, acylamino, amino substitutedacylamino, aryl, cycloalkyl, carboxy, alkoxy, aralkyloxy,alkoxycarbonyl, aralkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, (alkyl)(aralkyl)aminocarbonyl,aralkylaminocarbonyl, diaralkylaminocarbonyl, hydroxyl,carboxyalkylaminocarbonyl, hydroxylaminocarbonyl, mercapto, thioalkoxyor heterocycle; each R¹, independently, is selected from the groupconsisting of hydrogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, cycloalkylalkyl, aryl, aminoalkyl, mono ordi-alkyl-substituted aminoalkyl, mono- or di-aralkyl-substitutedaminoalkyl, hydroxyalkyl, alkoxyalkyl, mercaptoalkyl, andthioalkoxyalkyl; or R¹ and any A are taken together with the atoms towhich they are attached form a 3- to 6- membered ring heterocycle; eachR², independently, is selected from the group consisting of hydrogen andalkyl; n is an integer from 0 to 8; and X is selected from the groupconsisting of alkoxy, aryloxy, aralkyloxy, hydroxyl, amino, alkylaminooptionally substituted with hydroxy, aminocarbonyl,N-alkylaminocarbonyl, carboxy or alkoxycarbonyl, dialkylamino,cycloalkylamino, dicycloalkylamino, cycloalkylalkylamino,(alkyl)(aryl)amino, aralkylamino optionally substituted with carboxy,diaralkylamino, arylamino, heterocycle, (mono- or bis-carboxylicacid)-substituted alkylamine, heterocyclylamino, andheterocyclyl-substituted alkylamino; provided that the compound offormula I is expressly notN-carboxymethyl-N-(phenylacetyl-L-leucyl-L-aspartyl-Lphenylalanyl-L-prolyl)piperazine and expressly not phenylacetyl Lleucyl-L-aspartyl-L-phenylalanyl-D-proline amide; and provided that Z isnot pyridylcarbonyl, pyridylacetyl, phthalimido, 3-quinolinoyl,pyrazolylcarbonyl, or 2-pyrazinylcarbonyl, when Y² is D, Y³ is V, I, F,P, W, Y, or L, and n is 1; and also provided that Z is notpyridylcarbonyl, pyridylacetyl, phthalimido, 3-quinolinoyl,pyrazolylcarbonyl, or 2-pyrazinylcarbonyl, when both Y² is D and n is 0.2. The cell adhesion inhibitory compound according to claim 1, wherein:Z is selected from the group consisting of aliphatic acyl substitutedwith N-arylamido, heterocycloyl, arylsulfonyl, alkylsulfonyl,substituted aralkylcarbonyl, heterocycloalkylcarbonyl,cycloalkylcarbonyl optionally fused with aryl,heterocycloalkoxycarbonyl, alkylaminocarbonyl, arylaminocarbonyloptionally substituted with bis(alkylsulfonyl)amino,alkoxycarbonylamino, or alkenyl, and aralkylaminocarbonyl optionallysubstituted with bis-(alkylsulfonyl)amino, alkoxycarbonylamino, oralkenyl; each R¹, independently, is selected from the group consistingof hydrogen, alkyl, and aralkyl; and X is selected from the groupconsisting of alkoxy, aryloxy, aralkyloxy, hydroxyl, amino, alkylaminooptionally substituted with hydroxy, aminocarbonyl,N-alkylaminocarbonyl, carboxy or alkoxycarbonyl, dialkylamino,cycloalkylamino, dicycloalkylamino, cycloalkylalkylamino,(alkyl)(aryl)amino, aralkylamino optionally substituted with carboxy,diaralkylamino, arylamino, heterocycle, and (mono- or bis-carboxylicacid)-substituted alkylamine.
 3. The cell adhesion inhibitory compoundaccording to claim 1, wherein A¹ is cycloalkyl, alkyl optionallysubstituted with amino, acylamino, amino-substituted acylamino, aryl,carboxy, cycloalkyl, hydroxy, alkoxy, aralkyloxy, alkoxycarbonyl,aralkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, (alkyl)(aralkyl)aminocarbonyl,aralkylaminocarbonyl, diaralkylaminocarbonyl, alkoxycarbonylamino,mercapto, thioalkoxy, heterocycle, or A¹ and R¹ taken together withatoms to which they are attached form a heterocycle.
 4. The celladhesion inhibitory compound according to claim 3, wherein A¹ isaminocarbonylethyl, benzyl, n-butyl, isobutyl, carboxyethyl, cyclohexyl,1-hydroxyethyl, hydroxymethyl, mercaptomethyl, 1-methylpropyl,methylthioethyl, n-propyl, isopropyl, methoxycarbonylaminobutyl,6-aminohexanoylaminobutyl, or A¹ and R¹ taken together with atoms towhich they are attached form an azetidine, aziridine, pyrrolidine, orpiperidine.
 5. The cell adhesion inhibitory compound according to claim4, wherein A¹ is selected from the group consisting of benzyl, n-butyl,isobutyl, methylthioethyl, cyclohexyl, 1-methylpropyl, n-propyl andisopropyl.
 6. The cell adhesion inhibitory compound according to claim4, wherein A¹ and R¹ taken together with atoms to which they areattached form a pyrrolidine.
 7. The cell adhesion inhibitory compoundaccording to claim 1, wherein A² is alkyl optionally substituted withamino, aminocarbonyl, aryl, alkoxycarbonyl, aralkyloxycarbonyl,hydroxylaminocarbonyl, carboxy, NH-containing heterocycle, hydroxy,mercapto, aralkyl optionally substituted with amino, aminocarbonyl,carboxy, NH-containing heterocycle, hydroxy, mercapto, or A² and R¹taken together with atoms to which they are attached form a heterocycle.8. The cell adhesion inhibitory compound according to claim 7 wherein A²is carboxymethyl, 2-carboxyethyl, 1-carboxyethyl,hydroxylaminocarbonylmethyl, hydroxymethyl, mercaptomethyl,imidazolylmethyl, N-Bn-imidazolylmethyl, phenyl, carbomethoxymethyl,carbobenzyloxymethyl, or A² and R¹ taken together with atoms to whichthey are attached form an azetidine, aziridine, pyrrolidine orpiperidine.
 9. The cell adhesion inhibitory compound according to claim8 wherein A² is selected from the group consisting of carboxymethyl,2-carboxyethyl, 1-carboxyethyl, hydroxylaminocarbonylmethyl,hydroxymethyl, mercaptomethyl and imidazolylmethyl.
 10. The celladhesion inhibitory compound according to claim 1, wherein each A³,independently, is selected from the group consisting of amino acid sidechains and corresponding protected derivatives, cycloalkyl, and alkyloptionally substituted with aryl, cycloalkyl, carboxy,hydroxylaminocarbonyl, alkoxy, aralkyloxy, mercapto, N-containingheterocycle, carboxyalkylaminocarbonyl or amino-substituted acylaminogroup.
 11. The cell adhesion inhibitory compound according to claim 10,wherein each A³, independently, is selected from the group consisting ofamino acid side chains and corresponding protected derivatives,cyclohexyl, and alkyl optionally substituted with phenyl, cyclohexyl,carboxy, hydroxylaminocarbonyl, methoxy, benzyloxy, mercapto,N-benzylimidazolyl, biotinyl, tetrazolyl, valinyl-N-carbonyl or6-aminohexanoylamino.
 12. The cell adhesion inhibitory compoundaccording to claim 1, wherein each Y³, independently, is selected fromthe group consisting of amino acids and corresponding protectedderivatives of the amino acid.
 13. The cell adhesion inhibitory compoundaccording to claim 1, wherein: n is 2; Y¹ is leucinyl; Y² is aspartyl;and Y³ is valinylprolinyl.
 14. The cell adhesion inhibitory compoundaccording to claim 1, wherein X is selected from the group consisting ofalkoxy, aryloxy, aralkyloxy, hydroxyl, amino, mono- and dialkylaminooptionally substituted with hydroxy, aminocarbonyl,N-alkylaminocarbonyl, carboxy or alkoxycarbonyl, dialkylamino,cycloalkylamino, cycloalkylalkylamino, dicycloalkylamino,(alkyl)(aryl)amino, aralkylamino optionally substituted with carboxy,diaralkylamino, arylamino, N-containing heterocycle, bis-carboxylic acidsubstituted alkylamine, and (mono- orbis-carboxy)methylaminocarbonyl-substituted-N-containing heterocycle.15. The cell adhesion inhibitory compound according to claim 14, whereinX is selected from the group consisting of amino, methylamino,isopropylamino, isobutylamino, n-butylamino, t-butylamino, isoamyl,isopentylamino, hexylamino, cyclohexylamino, cyclohexylmethylamino,methylphenylamino, phenylmethylamino, phenylamino,4-methoxyphenylmethylamino, dimethylamino, diisopropylanino,diisobutylamino, hydroxy, methoxy, n-butoxy, t-butoxy, benzyloxy,2-piperidinecarboxylic acid,N′-(α,α′-bis-carboxymethyl)-2-piperidinecarboxamide,N′-carboxymethyl-2-piperidinecarboxamide,1-hydroxymethyl-2-methylpropylamino,1-N′-methylamido-1-methylethylamino, 3,3-dimethylbutylamino,1-N′-methylamidobutylamino, 1-amido-2-methylbutylamino,1-carbomethoxy-2-methylbutylamino, 1-N′-methylamido-2-methylbutylamino,1-carboxy-1-phenylmethylamino, morpholino, piperidinyl,N-phenylpiperazinyl, pipecolinyl, and piperazinyl.
 16. The cell adhesioninhibitory compound according to claim 1, wherein Z is selected from thegroup consisting of aliphatic acyl substituted with N-arylamido,substituted aralkylcarbonyl, heterocycloyl, andheterocycloalkylcarbonyl.
 17. The cell adhesion inhibitory compoundaccording to claim 2 selected from the group consisting of compound 1(L-Proline,1-[N-[N-[N-[[4-[[phenylamino]carbonyl]amino](3-methoxyphenyl)]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 2 (L-Proline,1-[N-[N-[N-[[4-[[phenylamino]carbonyl]amino](3-methoxyphenyl)]acetyl]-L-methionyl]-L-α-aspartyl]-L-valyl]-),compound 144 (L-Proline,1-[N-[N-[N-[[4-[[(2-hydroxy)phenylamino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 145 (L-Proline,1-[N-[N-[N-[[4-[[phenylamino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 146 (L-Proline,1-[N-[N-[N-[[4-[[(2-hydroxy)phenylamino]carbonyl]amino]phenyl]acetyl]-L-methionyl]-L-α-aspartyl]-L-valyl]-),compound 147 (L-Prolinamide,1-[N-[N-[N-[[4-[[phenylamino]carbonyl]amino](3-methoxyphenyl)]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 148 (L-Prolinamide,1-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 206 (L-Proline,1-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 315 (L-Proline,1-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-methionyl]-L-α-aspartyl]-L-valyl]-),compound 316 (L-Proline,1-[N-[N-[N-[[4-[[2-pyridylamino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 317 (L-Prolinamide,1-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino](3-methoxyphenyl)]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 337 (L-Serine,1-[N-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-L-prolyl]-),compound 338 (L-Threonine,1-[N-[N-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-L-prolyl]-L-seryl]-),compound 345 (L-Valine,1-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-),compound 346 (L-Proline,1-[N-[N-[N-[[4-[[(6-methyl-2-pyridyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 347 (L-Proline,1-[N-[N-[N-[[4-[[(2-fluorophenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 357 (L-Prolinamide,1-[N-[N-[N-[[4-[[(2-pyridyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 358 (L-Proline,1-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-,compound with 2 equivalents of2-amino-2-(hydroxymethyl)-1,3-propanediol), and compound 359 (L-Proline,1-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-,disodium salt).
 18. The cell adhesion inhibitory compound according toclaim 17 selected from the group consisting of compound 1 (L-Proline,1-[N-[N-[N-[[4-[[phenylamino]carbonyl]amino](3-methoxyphenyl)]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 206 (L-Proline,1-[N-[N-[N-[[4-[[(2-methylphenyl)ammo]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 316 (L-Proline,1-[N-[N-[N-[[4-[[2-pyridylamino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-),compound 358 (L-Proline,1-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-,compound with 2 equivalents of2-amino-2-(hydroxymethyl)-1,3-propanediol), and compound 359 (L-Proline,1-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-,disodium salt).
 19. The cell adhesion inhibitory compound according toclaim 18 is compound 358 (L-Proline,1-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-,compound 358 with 2 equivalents of2-amino-2-(hydroxymethyl)-1,3-propanediol).
 20. A pharmaceuticalcomposition comprising a compound according to claim 1 in an amounteffective for prevention, inhibition or suppression of cell adhesion anda pharmaceutically acceptable carrier.
 21. The pharmaceuticalcomposition according to claim 20, further comprising an agent selectedfrom the group consisting of corticosteroids, bronchodilators,antiasthmatics, antiinflammatories, antirheumatics, immunosuppressants,antimetabolites, immunomodulators, antipsoriatics, and antidiabetics.22. A cell adhesion inhibitory compound of formula (I),Z—(Y¹)—(Y²)—(Y³)_(n)—X  (I) or a pharmaceutically acceptable derivativethereof; wherein: Z is a (N—Ar′-urea)-para-substituted aralkylcarbonylgroup; Y¹ is —N(R¹)—C(R²)(A¹)—C(O)—; Y² is —N(R¹)—C(R²)(A²)—C(O)—; eachY³ is represented by the formula —N(R¹)—C(R²)(A³)—C(O)—; A¹ is selectedfrom the group consisting of amino acid side chains and correspondingprotected derivatives, cycloalkyl, and alkyl optionally substituted withamino, acylamino, amino substituted acylamino, alkoxycarbonylamino,aryl, cycloalkyl, carboxy, alkoxy, aralkyloxy, alkoxycarbonyl,aralkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, (alkyl)(aralkyl)aminocarbonyl,aralkylaminocarbonyl, diaralkylaminocarbonyl, hydroxyl,carboxyalkylaminocarbonyl, hydroxylaminocarbonyl, mercapto, thioalkoxyor heterocycle; A² is selected from the group consisting of acidicfunctional groups, and alkyl optionally substituted with an acidicfunctional group, protected acidic functional group or aryl; each A³,independently, is selected from the group consisting of amino acid sidechains and corresponding protected derivatives, aryl, cycloalkyl, andalkyl optionally substituted with amino, acylamino, amino-substitutedacylamino, aryl, cycloalkyl, carboxy, alkoxy, aralkyloxy,alkoxycarbonyl, aralkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, (alkyl)(aralkyl)aminocarbonyl,aralkylaminocarbonyl, diaralkylaminocarbonyl, hydroxyl,carboxyalkylaminocarbonyl, hydroxylaminocarbonyl, mercapto, thioalkoxyor heterocycle; each R¹, independently, is selected from the groupconsisting of hydrogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, cycloalkylalkyl, aryl, aminoalkyl, mono- or di-alkylsubstituted aminoalkyl, mono- or di-aralkyl-substituted aminoalkyl,hydroxyalkyl, alkoxyalkyl, mercaptoalkyl; and thioalkoxyalkyl, or R¹ andany A taken together with the atoms to which they are attached form a 3-to 6-membered ring heterocycle; each R² independently, is selected fromthe group consisting of hydrogen and alkyl; n is an integer from 0 to 8;and X is selected from the group consisting of alkoxy, aryloxy,aralkyloxy, hydroxyl, amino, alkylamino optionally substituted withhydroxy, aminocarbonyl, N-alkylaminocarbonyl, carboxy or alkoxycarbonyl,dialkylamino, cycloalkylamino, dicycloalkylamino, cycloalkylalkylamino,(alkyl)(aryl)amino, aralkylamino optionally substituted with carboxy,diaralkylamino, arylamino, heterocycle, (mono or bis-carboxylicacid)substituted alkylamine, heterocyclylamino, andheterocyclyl-substituted alkylamino.
 23. The cell adhesion inhibitorycompound according to claim 22, wherein each Y³ is independentlyselected from the group consisting of amino acids and correspondingprotected derivatives of the amino acids.
 24. The cell adhesioninhibitory compound according to claim 22, wherein: Y¹ is leucinyl; Y²is aspartyl; and Y³ is valinylprolinyl.
 25. The cell adhesion inhibitorycompound according to claim 22, wherein Z is a(N—Ar′-urea)-para-substituted-phenylmethylcarbonyl.
 26. The celladhesion inhibitory compound according to claim 22, wherein: Z is a(N—Ar′-urea)-para-substituted aralkylcarbonyl group; each R¹,independently, is selected from the group consisting of hydrogen, alkyl,and aralkyl; and X is selected from the group consisting of alkoxy,aryloxy, aralkyloxy, hydroxyl, amino, alkylamino optionally substitutedwith hydroxy, aminocarbonyl, N-alkylaminocarbonyl, carboxy oralkoxycarbonyl, dialkylamino, cycloalkylamino, dicycloalkylamino,cycloalkylalkylamino, (alkyl)(aryl)amino, aralkylamino optionallysubstituted with carboxy, diaralkylamino, arylamino, heterocycle, and(mono- or bis-carboxylic acid)-substituted alkylamine.
 27. The celladhesion inhibitory compound according to claim 22, wherein A¹ iscycloalkyl, alkyl optionally substituted with amino, acylamino,amino-substituted acylamino, aryl, carboxy, cycloalkyl, hydroxy, alkoxy,aralkyloxy, alkoxycarbonyl, aralkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, (alkyl)(aralkyl)aminocarbonyl,aralkylaminocarbonyl, diaralkylaminocarbonyl, alkoxycarbonylamino,mercapto, thioalkoxy, or heterocycle, or A and R′ taken together withatoms to which they are attached form a heterocycle.
 28. The celladhesion inhibitory compound according to claim 27, wherein A¹ isaminocarbonylethyl benzyl, n-butyl, isobutyl, carboxyethyl, cyclohexyl,1-hydroxyethyl, hydroxymethyl, mercaptomethyl, 1-methylpropyl,methylthioethyl, n-propyl, isopropyl, methoxycarbonylaminobutyl,6-aminohexanoylaminobutyl, or A¹ and R¹ taken together with atoms towhich they are attached form an azetidine, aziridine, pyrrolidine, orpiperidine.
 29. The cell adhesion inhibitory compound according to claim28, wherein A¹ is selected from the group consisting of benzyl, n-butyl,isobutyl, methylthioethyl, cyclohexyl, 1-methylpropyl, n-propyl, andisopropyl.
 30. The cell adhesion inhibitory compound according to claim28, wherein A¹ and R¹ taken together with atoms to which they areattached form a pyrrolidine.
 31. The cell adhesion inhibitory compoundaccording to claim 22, wherein A² is alkyl optionally substituted withamino, aminocarbonyl, aryl, alkoxycarbonyl, aralkyloxycarbonyl,hydroxylaminocarbonyl, carboxy, NH-containing heterocycle, hydroxy,mercapto, aralkyl optionally substituted with amino, aminocarbonyl,carboxy, NH-containing heterocycle, hydroxy, mercapto, or A² and R¹taken together with atoms to which they are attached form a heterocycle.32. The cell adhesion inhibitory compound according to claim 31, whereinA² is carboxymethyl, 2-carboxyethyl, 1-carboxyethyl,hydroxylaminocarbonylmethyl, hydroxymethyl, mercaptomethyl,imidazolylmethyl, N-Bn-imidazolylmethyl, phenyl, carbomethoxymethyl,carbobenzyloxymethyl, or A² and R¹ taken together with atoms to whichthey are attached form an azetidine, aziridine, pyrrolidine orpiperidine.
 33. The cell adhesion inhibitory compound according to claim32, wherein A² is selected from the group consisting of carboxymethyl,2-carboxyethyl, 1-carboxyethyl, hydroxylaminocarbonylmethyl,hydroxymethyl, mercaptomethyl, and imidazolylmethyl.
 34. The celladhesion inhibitory compound according to claim 22, wherein A³ is,independently, selected from the group consisting of amino acid sidechains and corresponding protected derivatives; cycloalkyl; and alkyloptionally substituted with aryl, cycloalkyl, carboxy,hydroxylaminocarbonyl, alkoxy, aralkyloxy, mercapto, N-containingheterocycle, carboxyalkylaminocarbonyl, or amino-substituted acylaminogroup.
 35. The cell adhesion inhibitory compound according to claim 34,wherein A³ is independently selected from the group consisting of aminoacid side chains and corresponding protected derivatives, cyclohexyl,and alkyl optionally substituted with phenyl, cyclohexyl, carboxy,hydroxylaminocarbonyl, methoxy, benzyloxy, mercapto, N-benzylimidazolyl,biotinyl, tetrazolyl, valinyl-N-carbonyl, or 6-aminohexanoylamino group.36. A pharmaceutical composition comprising a compound according toclaim 22 in an amount effective for prevention, inhibition orsuppression of cell adhesion and a pharmaceutically acceptable carrier.37. The pharmaceutical composition according to claim 36 furthercomprising an agent selected from the group consisting ofcorticosteroids, bronchodilators, antiasthmatics, antiinflammatories,antirheumatics, immunosuppressants, antimetabolites, immunomodulators,antipsoriatics and antidiabetics.
 38. The cell adhesion inhibitorycompound according to claim 18 is compound 359 (L-Proline,1-[N-[N-[N-[[4-[[(2-methylphenyl)amino]carbonyl]amino]phenyl]acetyl]-L-leucyl]-L-α-aspartyl]-L-valyl]-,disodium salt).